Human Cheek Cells

Fresh Mount slidefree-hand sections cut with a razor blade of part or

all of an organism, or material which is mechanically or chemically
teased apart. The material is placed in a drop of physiological
saline or water in the center of a clean slide. Some care must be
taken in lowering the cover slip in order to avoid trapping air
bubbles. The difficulty is overcome by first allowing only one edge
of the cover slip to rest on the slide and, while supporting the
cover slip with a needle, slowly lowering it over the preparation.
Since this type of preparation will dry up, a drop of saline or water
must occasionally be added along one edge of the cover slip.
Alternatively you can reduce evaporation by ringing the edges of
the cover slip with a thin film of vaseline. It is often helpful to
increase the contrast of certain cell structures so that they can be
seen more clearly. Chemicals that dye parts of cells for this
purpose are called stains. A great variety of stains are available;
some colour all parts of cells more or less indiscriminately, while
others act more specifically on particular structures or chemical
compounds within the cell. Section F. Cheek Squamous Epithelial
Cells
Magnified view (400x) of squamous epithelial cells from the buccal mucosa
(cheek cells from inside the mouth). The cells are stained with a dye called
methylene blue. The nucleus and cell membrane are clearly visible. Plant
structures such as a cell wall, chloroplasts and large central vacuole are absent.
Because they don't have a rigid (firm) cellulose cell wall, these cells are flimsy
and irregular in shape, unlike the rectangular shape of the onion cells. Although
they were photographed in fall of 2001, these cells actually came from a student
in a previous biology lab three years before.
Name
The Human Cheek Cell _______________________________
___
1. List the 3 parts of the Cell Theory Procedure:
1. Put a drop of methylene blue on a
_____________________________________ slide. Caution: methylene blue will stain
_____________________________________ clothes and skin.
2. Gently scrape the inside of your cheek
_____________________________________ with the flat side of a toothpick. Scrape
lightly.
2. Describe or define each of the following 3. Stir the end of the toothpick in the stain
--cell and throw the toothpick away.
membrane ____________________________ 4. Place a coverslip onto the slide
5. Use the SCANNING objective to focus.
_ You probably will not see the cells at this
--cytoplasm ___________________________ power.
______ 6. Switch to low power. Cells should be
visible, but they will be small and look like
--nucleus nearly clear purplish blobs. If you are
___________________________________ looking at something very dark purple, it
is probably not a cell
7. Once you think you have located a
--organelle ____________________________ cell, switch to high power and refocus.
____ (Remember, do NOT use the coarse
adjustment knob at this point)

3. Sketch the cell at low and high power. Label the nucleus, cytoplasm, and cell
membrane. Draw your cells to scale.
Low Power High Power

5. Why is methylene blue necessary?

6. The light microscope used in the lab is not powerful enough to view other organelles
in the cheek cell. What parts of the cell were visible.

6. List 2 organelles that were NOT visible but should have been in the cheek cell.
7. Is the cheek cell a eukaryote or prokaryote? How do you know?
8. Keeping in mind that the mouth is the first site of chemical digestion in a human.
Your saliva starts the process of breaking down the food you eat. Keeping this in mind,
what organelle do you think would be numerous inside the cells of your mouth?
Cheek - Cheeks ( Latin: buccae) constitute the area of the face below the eyes and between the nose
and the left or right ear. It is fleshy in humans and other mammals, the skin being suspended by the
chin and the jaws, and forming the lateral wall of the human mouth, visibly touching the cheekbone..

Methylene blue

Methylene blue is used to stain animal cells, such as human cheek cells, to make their nuclei more
observable. Also used to staining the blood film and used in cytology.

Section D. Potato Tubers & Banana Fruits

T he thin-walled parenchyma cells of a potato tuber are filled with membrane-

bound, starch-storage organelles called amyloplasts. They are also referred to as
"starch grains" in most general biology textbooks. Since iodine stain (gram's
iodine) makes starch turn purplish-black, the amyloplasts can easily be viewed
with a compound microscope (400x). Insoluble starch (amylopectin) is deposited
in concentric layers within the amyloplasts. Unlike the long, coiled molecules of
soluble starch (amylose), the molecules of amylopectin are much shorter, with
only 40-60 glucose subunits. Amylopectin molecules consist of highly branched
chains that do not coil. Starch grains of different plant species have characteristic
shapes, such as maize (corn), oats, bananas, potatoes and wheat. For example,
banana starch grains are more elongate than potato starch grains. Starch is
hydrolyzed (broken down) by amylase enzymes (including B-amylase and
maltase). During hydrolysis a water molecule is inserted between each glucose
subunit. Starch is typically stored in underground organs, including storage roots,
rhizomes, tubers, corms and bulbs.
Magnified view (400x) of several parenchyma
cells of a potato tuber showing the thin,
transparent cell walls and clusters of amyloplasts
(starch grains). The starch grains were stained
black with gram's iodine. Iodine is used in chemistry as an indicator
for starch. When starch is mixed with iodine in solution, an intensely dark blue color develops,
representing a starch/iodine complex. Starch is a substance common to most plant cells and so a
weak iodine solution will stain starch present in the cells. Iodine is one component in the staining
technique known as Gram staining, used in microbiology. Lugol's solution or Lugol's iodine (IKI) is a
brown solution that turns black in the presence of starches and can be used as a cell stain, making the
cell nucleimore visible.Iodine is also used as a mordant in Gram's staining, it enhances dye to enter
through the pore present in the cell wall/ cell membrane.

Paramecium
• Obtain a prepared slide of paramecium and view it using the scanning lens
(40X). After the light and focus are adjusted, center a paramecium and
increase the magnification to 100X. Next, adjust the lighting, use the fine
focus, and then center the paramecium. Increase the magnification to 400X
and then adjust the light and adjust the fine focus.
• The procedure described above (adjust the light, adjust the focus, center the
specimen, increase the magnification) should be used whenever you are
trying to view a small specimen that is difficult to find.
7) Draw the cell. Label the cilia and the oral groove.
Below: Paramecium.
Click on the
image to view
an enlarge
ment.

The beating of the cilia is easily visible under light microscopic examination,
especially if the movement of the organism has been retarded by the addition of a
viscous compound such as glycerol to the sample.

In this photo two layers of onion cells are visible. The cells
in the top layer are round and the cells in the
bottom layer are rectangle in shape. (View enlarged)
 The Structure of a Plant Cell

To depict the structure of a plant cell, an epidermal cell of an onion will be used as
an example. The epidermis is the final tissue that covers all organs above ground.
The cells of the onion epidermis are common specimens on the first day of a
German basic botanical course. Since they contain no chlorophyll, they are actually
no "typical" plant cells.
The picture above shows an onion"s epidermal cells. They are elongated and the
ratio of length to width can vary strongly. Each cell is enclosed by a wall. In the
region of the cell poles and where three cells adjoin, large intercellular spaces can
be observed. Elsewhere a pectin-containingmiddle lamina cements neighboring
cells together like bricks. The cell wall is perforated at regular intervalls, so that
adjoining cells are in contact. The holes of the perforation are called simple
pits and the plasma cords that run through them plasmodesmata . The surface of
the epidermal cells seems to be folded, an effect that is caused by the water-
repellent, waxy cuticle.

Protoplasm, Cytoplasm and Cytosol: The Cell's Content

The "living" content of a cell, the protoplasm , is surrounded by a membrane
called plasmamembrane or plasmalemma. The protoplasm is usually next to the
cell wall, so that the plasmalemma can hardly be seen. To display it, the cells are
transferred into a high salt or sugar solution. As a result the protoplasm shrinks and
detaches itself from the wall. The process is reversible and is called plasmolysis.
This behavior is due to the properties of the membrane and the plasma. It is
reviewed in more detail elsewhere. A substance that causes plasmolysis is
called plasmolyticum and - depending on its chemical composition (potassium
ions orcalcium ions, for example) - the protoplasm takes on different shapes. The
plasmolyticum has accordingly an influence on the properties of the membrane.
The properties of the plasmamembrane differ from that of the tonoplast. The
tonoplast is the membrane that surrounds the vacuole. The difference is especially
striking if cells with a colored vacuole content are used. Often the vacuole is criss-
crossed by numerous plasma cords. The plasma cannot therefore not simply be
viewed as a solution that is influenced by the rules of hydrodynamics alone. Rather,
it contains viscous, structure-determining components, whose chemical,
physicochemical and structural properties have only been recognized recently and
in fragments.

Cytoplasm and Caryoplasm

The nucleus is a rather conspicuous part of nearly every living plant cell. Its
structure separated from the rest of the cell by the nuclear envelope, a membrane
system that consists of two discrete membranes as can be seen on
electromicroscopic images. The nuclear content is called the caryoplasm while that
of the rest of the cell is called cytoplasm. But these terms are only valid at certain
stages of a cell's life cycle. In the course of cell-division and mitosis, the nuclear
envelope disintegrates and the nucleus is replaced by the chromosomes. It makes
consequently no sense to speak of caryo- and cytoplasm during these stages.
The nucleus of plant cells is usually of a round or elliptic appearance, sometimes it
is also shaped like a spindle. One nucleus per cell is the rule, but cells with two or
more nuclei are no rare exception. The cells of certain algae of the
genus Chladophora have many nuclei, they are polyenergid. The nucleoli that can
often be perceived after staining are substructures of the nucleus. They, too,
disintegrate during cell division and mitosis and do not reshape before a new
nucleus has been formed.

Plastids

Plastids are organelles that occur only in plants. Their most prominent members are
the chloroplasts. Others plastids are the colored chromoplasts and the colorless
leucoplasts as well as their proplastids. Proplastids are vestigial bodies that are
generated during germ cell development due to degeneration of plastids, for
example. They may differentiate into complete plastids during the development of
the plant embryo. Their ripening into chloroplasts occurs usually only after light
exposure.
 © M. Knee

Chloroplasts contain the green plant color chlorophyll. They are the places
where photosynthesis takes place. Chloroplasts enable the plants to convert solar
energy into chemical energy. Because of this process, plants are called primary
producers. The existence of consumers, like most animals, depends on them.
Chloroplasts occur in most cell types, but only in organs above ground. They can
be especially well observed in tissues consisting of a single layer as in the leaflike
structures of some mosses (Funaria hygrometrica or Mnium hornum) or in the
water plant Vallisneria). Here they are rather large and of a lens-shaped
appearance. During daytime, when the light is diffuse, they occur mainly at the
upper and lower surface of the chloroplast. They appear to be round under top
view. If exposed to strong light, they gather in parallel to the lateral sides of the cell
which gives them an elleptic appearance upon top view.
Chloroplasts are the site of starch production and -storage. Starch can easily be
detected with the aid of potassium iodide (LUGOL's reagent). The starch-iodine
complex is deeply blue-violet.
Starch production during photosynthesis can be made visible by placing a mask at a
leaf that covers it partially while leaving some places exposed to sunlight. After one
day of exposure, the leaf is first bleached to get rid of other pigments and
afterwards treated with potassium iodide. An image is gained that is the exact
replication of the mask and at the same time represents starch synthesis in the leaf.
This experiment has first been done by J. v SACHS, probably the most outstanding
plant physiologist of the 20th century. He thought that starch was the primary
product of photosynthesis. This assumption proved wrong. It is well-known today
that the first products of photosynthesis are monomeric sugars (glucose and others)
and that only part of them is used for starch production.
The structures of the chloroplasts of higher cells resemble largely that of mosses.
Their average diameter is 4 - 8 mm, an average cells contains 10 through 50. Their
chlorophyll is unevenly distributed. At high resolutions chlorophyll-rich and
chlorophyll-poor areas can be distinguished. This is due to the inner structure of the
chloroplast: it is organized into grana (chlorophyll-rich) and stroma (chlorophyll-
poor). Upon stimulation with short-waved light (blue or violet) the chlorophyll
emits an intensive red autofluorescence that looks especially impressive in a
fluorescence microscope. The differences between grana and stroma become very
obvious. The uniformity of the chloroplasts of all higher plants points out that the
optimal form has been found rather early in evolution and has not been changed
since. This is different with algae. The chloroplasts of green algae
(Chlorophyceens) are very varied in shape. Many species have just one chloroplast
that covers nearly the whole space of the cell's interior. It is screw-like
in Spirogyra-species, star-shaped in Zygnema and Zygnemopsis and netlike
in Oedogonium.
The disc-shaped chloroplast of Mougeotia can be viewed either from above or in
profile depending on the amount of light used. Its rotation is a well-analyzed
example of an induced chloroplast movement. The chloroplasts of many species of
algae contain often well-visible pyrenoids, structures, that produce and structure
starch.
Chromoplasts are red, orange or yellow plastids. The color is usually the result of
yellow xantophyll and yellow to red carotinoids. Both compounds do also exist in
chloroplasts, but are concealed by chlorophyll. Chlorophyll is broken down much
faster than carotinoids as can be observed in the colored leaves in autumn. Fluid
transitions between chromo- and chloroplasts exist, just as between chromo- and
leucoplasts. Typical chromoplasts cause the orange color of the carrot, the red color
of the ripe pimento and tomato as well as the color of numerous flowers.
Carotinoids are not very water-soluble and do therefore often crystallize within the
chromoplasts. Their crystals can be disc-shaped, needle-like, jagged or sickle-like.
In many cases, flower and leaf colors are caused by the colored content of the
vacuole. The color of the vacuole and that of the plastids may lead to a mixed color.
The leaves of the copper beech, where the vacuole's content is red and that of the
chloroplasts is green are a typical example. The plastids of the red and brown algae
are traditionally counted among the chromoplasts although they contain
chlorophyll. The green colour is concealed by the red phycoerythrin
(Rhodophyceae) or the brown fucoxanthin (Phaeophyta).
Leucoplasts are common, colorless plastids. They develop from proplastids, but
form no homogeneous group of their own. A part of them can differentiate into
chloroplasts or chromoplasts at light exposure, while this is not due for others.
The guard cells, for example, contain leucoplasts, that are permanently exposed to
light without developing into chloroplasts. Leucoplasts do also occur within
colorless leaves (variegated leaves) or plant parts. There exists a number of
examples which show that they developed from chloroplasts that lost their ability to
produce chlorophyll. There are even species, like Neottia, an orchid that cannot
produce chlorophyll at all and are thus dependent on a parasitic or saprophytic
lifestyle (saprophy is the feeding from dead organic material).
A second class of leucoplasts occurs within the non-green tissues of otherwise
green plants. It is especially common within roots. Though these leucoplasts are
capable to become green, they do usually not since they are not exposed to light.
The leucoplasts of the calyptra (a calyptra is any hood or cap of cells protecting a
plant part) contain starch and are therefore counted among the amyloplasts (starch-
containing leucoplasts). They have, as is explained later, the function of statolithes,
that have an important part in the perception of gravity (geotropism).

Starch
We got to know starch in the section above as a content of chloroplasts and
leucoplasts (amyloplasts). It is produced by the polymerization of glucose residues,
which again are products of photosynthesis. Since the plant is able
to transport sugars from leaf to root or from leaf to seed and fruit, starch production
can also take place in these organs. Different species produce starch grains of
different shape. Since the shape of starch grains informs about their origin, they are
helpful in the identification of seeds and other starch-containing plant parts. The
following numbers show the variations in their diameters. Starch grains from potato
tubershave a diameter of 70 - 100 µm, that of the endosperm of wheat 30 - 45 µm
and that of corn endosperm 12 - 18 µm. Their shape reflects their development.
Thestarch molecules are long-stretched and only sparsely branched. They are
deposited within the plastids and their development begins at a so-called formation
center from where it proceeds radially. Layer follows layer and the thickness of one
layer is dependent on the average molecular length. A starch grain is therefore
organized like a crystal (semi-crystalline). This can be shown very impressively
with a polarization microscope. Within the moistened specimen can a layering be
viewed that is dependent on the water content of the single molecular parts.
A. Model of a
starch grain
structure. The
single lines
symbolize starch
molecules. They
are arranged in a
radial
pattern. B. Layerin
g of the starch
grains. a.
Formation center
and layer borders, b. Diagram of the refraction conditions. The ordinate shows the
refraction index. (according to A. FREY-WYSSLING, 1938)
The denser the molecular packaging, the less water is deposited. Layers with less
water content refract light stronger than those with much water. After drying of the
preparation no layering can be perceived any more. Depending the central or
peripheral placing of the formation center, starch grains with either concentric or
eccentric layering develop. The starch grains of graminaceous plants (wheat, corn,
etc.) are usually concentric, while those of the potato are always eccentric.
Sometimes plastids with two to three formation centres occur in graminaceous
plants. This leads to the development of several starch grains. During growth
common outer layers may be formed.
Compound starch grains are typical for oat, they are built from a large number of
smaller grains.
Starch grains in bean seeds (Phaseolus vulgaris) are very big, their shape is round
or oval, the spacing of the layers is very regular. Their centers can easily be
hollowed out by addition of water displaying radial ruptures in microscopic images.
In the sap of Euphorbia splendens dumb-bell shaped starch grains can be found.

Crystalline Inclusions within Cells

Many plant cells contain crystalline inclusions of

different chemical composition and shape.
Crystalline aggregations are called druses,
bundles of needle-shaped crystals are termed
raphids. Scanning electron microscopic
images: Top picture: Calcium oxalate druse in
the mesophyll cells of an oleander leaf (Nerium
oleander). Typical druse shape of dicots. Middle
picture: Calcium oxalate needles (raphids) of a
vanilla root (Orchidaceae). Typical raphid
bundle of monocots. Lowest picture: Silicate
bodies of silicate cells in the epidermis
of Schizachyrium sanguineum (a gramineaen
species of the old world tropics). Characteristic
mineralization of a gramineaen cell. [W.
BARTHLOTT, MARTENS, 1979 (c), W.
BARTHLOTT, unpublished (a, b)]

The Cell Wall

Except for very few examples, plant cells are surrounded by
a cellulose containing cell wall. It is flexible and distortable during growth, but
loses its ability for distortion after growth has stopped, while a limited flexibility
remains. Because of these changes, it is distinguished between primary and
secondary cell walls. As we will see when talking about electron microscopic
pictures of the cell wall, both forms differ mainly in the arrangement of their
cellulose microfibrils. While they are unorganized within the amorphous matrix of
the primary cell wall, they are organized into several ordered layers that are
arranged one on top of the other at right angles in the secondary cell wall. The
secondary cell walls of many cells, especially those of vascular tissues, are
incrusted with strengthening material. Two important ones are:
lignin, the ground substance of wood and
suberin, the ground substance of cork
Their details are reviewed here. In addition, secondary walls contain often phenolic
oxidation products that lends them a dark color (red to black with various shades).
CELLS and the LIGHT MICROSCOPE
OBJECTIVES
1. Understand the theory and use of the compound microscope
2. Distinguish the difference between magnification and
resolution.
3. View and understand the structures of a typical plant and
animal cell.
4. Locate and identify the organelles that are visible with the
light microscope.

This laboratory is to help you to understand the function of the bright field
microscope and introduce you to the world of microscopic anatomy - studying
biology from the perspective of the individual cell. The cell is the basic unit of life.
Some cells are free-living, others become more highly specialized and form
integrated aggregates, associating together to form more complex organisms. There
are cells that look rather "simple" inside, others appear much "busier" and more
complex. Some cells have plant-like characteristics, others are animal-like. There is
no such thing as a "typical" cell which incorporates and displays all the various
internal and external structures. One must examine a number of different cell types
to appreciate the great diversity, and the great similarity of form.
Once you have familiarized yourself with the basic operation of the light
microscope, review the basic organization of different cells and get a feel for how
the cell has become adapted to a variety of biological roles.
I. THE LIGHT MICROSCOPE
For well over a century, the light microscope has been one of the most important
instruments available to the biologist. This instrument is used to extend the range of
our vision. Ordinarily, humans cannot see objects smaller than 0.1 mm in diameter,
but the light microscope renders objects as small as 0.2 microns (1 micron = 10-
3
mm) visible to our eye. The modern compound microscope is a precision
instrument, designed to perform particular functions in a particular way. When the
microscope is used correctly, it will disclose many structures that the beginning
student will probably not see. For this reason, do not be surprised or skeptical if at
first you fail to see many of the details in plant or animal cell. How much you see
depends mostly on how well you adjust the instrument. Below are outlined some of
the various parts of the instrument (see fig. 1-1) and their function, as well as brief
instructions on how to use the instrument to achieve maximal results.
A. Terms Used in Microscopy
Magnification An image being seen at a magnification of 100X
means that its linear dimensions are 100 times those of the object
giving rise to the image. Thus, magnification is the ratio of the
apparent size of the object as seen through the microscope
(image size) to the real size of the object (object size). It is
expressed in units known as diameters, abbreviated as X (e.g.,
100X). The compound microscope has two separate lens systems,
the objective and the ocular. The objective, nearest the specimen,
magnifies the specimen a certain amount, and the ocular further
magnifies this image. Thus the image as seen by the eye has a
magnification equal to the product of the magnifications of the
two systems.

Resolution This refers to the ability of the microscope to distinguish as separate

and distinct objects that lie in close proximity. Magnification is not the sole aim of
a microscope. There is no upward limit to the magnification of a microscope, but
there is a limit to useful magnification. One can have increase in magnification
without an increase in visible detail. The basic limit in seeing objects is resolving
power, not magnification, and the physical phenomenon of diffraction of light
imposes limits on the resolving power. Attaining the maximum resolution depends
primarily on the design of the objective lens (see numerical aperture below). An
objective lens capable of utilizing a large angular cone of light coming from the
specimen will have a higher resolving power than a lens limited to a smaller cone
of light. Of course the resolvable detail must be magnified a sufficient amount in
order to be seen - hence the relationship between magnification and resolution.
Numerical Aperture This is a measure of the angle of the maximum cone of light
that can enter the objective lens. It is expressed by the equation: N.A. = n (sin q),
where n is the index of refraction of the medium between the object and the
objective and q is one half the angle of the entering cone of illumination. The
greater the N.A., the greater the resolving power.
Working distance This is the distance between the front of the objective, when it
is focused on the specimen, and the specimen itself. When viewing thin
preparations mounted on microscope slides and covered with a cover glass, it is the
distance to the top of the cover glass.
Field of view The circular field you see when you look through the ocular lens.
The field of view changes in size at different magnifications.
Depth of focus This refers to the thickness of the specimen which may be seen in
focus at any one time. As you focus up and down on your specimen, you will
notice that only a thin layer is in focus, not the entire thickness. The greater the
N.A. and magnification, the thinner is the layer in focus at any one time.
B. Mechanical Components
1. The mechanical stage is the square, horizontal platform that serves as a shelf to
support the material to be examined. The hole in the center of the stage serves to let
the light from the illuminator and condenser shine upon the specimen. On the
surface of the stage there is a special device to securely hold the specimen (i.e., the
microscope slide) and, using the two knobs below the right side of the stage, one
can move the specimen left and right as well as front and back.
2. There are two focusing knobs found on either side of the support arm. The larger
ones are the coarse adjustment and the smaller are for fine focusing. Both of these
move the stage up or down, depending on the direction turned, and serve to change
the distance between the specimen and the objective lens.

Figure 1-1 Typical Binocular Compound Light Microscope

C. Optical Components
1. The light source is located in the base of the microscope, and the light beam
passes out through the illuminator lens. Light is controlled by a rotating switch
located on top of the microscope base.
2. The light passes into the condenser, which concentrates the light onto the
specimen. On the left side of the instrument, beneath the stage, is the condenser
focusing knob, and it controls the position of the condenser relative to the
specimen. Associated with the condenser is the iris diaphragm, which in your
microscope serves to control the amount of illumination reaching the specimen.
There is a lever which, when moved back and forth, serves to open and close the
iris diaphragm, altering the amount of light passing through. A very common fault
in microscopy is to close down the iris diaphragm too far; contrast improves, but
there is considerable loss of resolution and other undesirable image artifacts are
introduced. You will be instructed in the proper adjustment of this critical
component.
3. Immediately above the specimen is the revolving turret which carries the
objective lenses. These form the initial magnified image of the specimen and
consists basically of a series of lenses of different sizes. There is the "scanning"
objective which magnifies 4 times, a low power objective that magnifies 10 times,
and a high power objective that magnifies 40 times; the oil immersion objective
(probably will not be used in this course) magnifies 100 times. One begins all
observations with the lower power objectives and works up to the high power one.
They are changed by rotating the turret, usually in a clockwise direction.
4. The lens system next to the eye is called the ocular, and this further magnifies
the image and projects it onto the retina of your eye. The ocular lenses (fitted
loosely into your microscope) have a magnifying power of 10X. In a binocular
microscope, the two ocular lenses can be adjusted to accommodate the distance
between the eyes and their individual focusing.
D. Preparation for Viewing
1. Clean the lenses. Dust and dirt will accumulate on the ocular lens, therefore, each
time you use a microscope, clean the lenses with lens paper. Also clean the
objective lens since other students use the same instrument and may cause water
and dirt to soil the objective lenses.
2. Make sure the 4X or 10X objective is in place. Raise the condenser to its upper
position. Turn on the illuminator.
3.Place your slide on the stage with the specimen over the condenser lens. Focus
the specimen with the coarse focus knobs (the larger ones).
4.With the specimen in focus, lower the condenser until the illuminator is in focus.
You will see sharp images of the lint on top of the illuminator. Then move the
condenser down very slightly to even out the illumination.
5.Remove the right ocular lens and look down the tube, keeping your eye several
inches away. Open and close the iris diaphragm and notice its image down by the
objective lens. Adjust this so that almost the entire area of the objective lens is
illuminated; this provides optimum definition and resolution for the available
lighting. If you close it down excessively, you can see there is increased contrast,
but this will reduce resolution and image quality. Try it and see.
6. If you want higher magnification and resolution, swing in the high power
objective. The objective lenses are parfocal, meaning the specimen is close to focus
from one objective to another. You will need to turn the fine focus knobs slightly.
You should not need to use the coarse focus. When you go from the scanning lens
to the 10X lens, you may need to use the coarse focus knobs.
7. You must repeat step 5 each time you change objectives!
E. Physical Properties of the Objective Lenses
1. Examine each objective lens and record its magnification and numerical aperture
in the table below.
2. Measure the diameter of the field of view of the 10X and 40X lenses by counting
the number of microns seen in a stage micrometer, and record the result in the table
below. Can you detect any relationship between magnification and field of view?
3. Obtain a slide with three colored threads that have been mounted one on top of
another. Use your microscope to determine which color thread is on top and which
is on the bottom. TOP: _____________ BOTTOM: _____________.

Common Name Working Magnificati N.A.

Distance on

SCANNING LENS 25 - 55 mm

LOW POWER 5 - 10 mm

HIGH POWER 0.15 - 0.6 mm

OIL IMMERSION 0.05 - 0.15

mm

F. Microscopic Dimensions
The metric system is used exclusively in science for making and reporting
measurements of weight, volume and distance. The table below explains the
various units used in microscopic measurements.

Definition Description

meter (m) =1,000 millimeters, 9.37 inches

millimeter (mm) =1,000 µm , or the smallest unit on your ruler

micron or micrometer =1,000 nm; animal cells average 50 microns in

(µm) diam.; the resolving power of the light
microscope is 0.2 µm.

nanometer (nm) =10 Angstroms, used to measure size of large

molecules resolving power of electron microscope
is 0.1 nm

II. EUKARYOTIC PLANT-LIKE CELLS

A. Onion Epidermis
1. An onion scale is lined on either side by a single layer of cells called
an epidermis. Obtain a piece of onion epidermis by bending a piece of onion until it
snaps. The two halves will only be attached by the epidermis. Carefully peel off the
transparent epidermis from the rest of the onion piece.
2. Lay the peel flat on a clean microscope slide in a drop of acetocarmine, making
sure the tissue isn't folded back on itself. Add a coverslip and view under the low
power objective.
3. Not all the organelles will be clearly visible. You should see the cell shape, the
large central vacuole which is colorless, and the nucleus. Why don't you see the
plasma membrane? How about mitochondria, or ribosomes?
4.The nucleus will stain reddish and the cytoplasm will be pink. You should be able
to distinguish between the vacuole and cytoplasm. Where is the nucleus located
within the cell? Can you see the plasma membrane now?

B. Elodea Leaf
1. Make a wet mount of a small leaf taken from near the tip of an Elodea shoot,
placing the upper surface of the leaf uppermost on the slide.
2. Focus the leaf using low power, find a portion where the cells are clearly visible
and switch to higher power. Due to the thickness of the cells, you will have to
continually focus up and down to see all the parts. This is an example of the
concept of the depth of focus. How many cell layers thick is the leaf?
3. The cells of Elodea are considerably more complex than the onion epidermal
cells. You should be able to see the cell wall; and inside the cell wall is the
cytoplasm, an almost transparent, colorless substance. The nucleus, with a
nucleolus, is difficult to see, but it is located near the periphery of the cell. The
vacuole occupies the center of the cell. All of the cell membranes are below the
resolving power of the light microscope, as is the plasma membrane. The most
obvious structures are thechloroplasts, small green bodies located in the
cytoplasm.
4. Look closely at the chloroplasts in cells located near the mid-vein of the leaf.
You should find cells in which the chloroplasts are moving around the perimeter of
the cells. This movement is the result of cytoplasmic streaming or cyclosis. Are the
chloroplasts in the same cell all moving in the same direction? Are the chloroplasts
within different cells moving in the same direction?
5. Draw and label a typical Elodea cell as it appears under high power.

III. EUKARYOTIC CELLS

A.Amoeba
The Amoeba is a free-living, single cell, and all of the functions and activities we
equate with the living state are carried out within this single cell. You may have
heard these organism being referred to as "primitive", but after you examine
the Amoeba and Paramecium, you may wonder what that term means when used in
reference to unicellular organisms.
1. Using the eye dropper, obtain a small sample of the detritus from the bottom of
the culture jar, place it on the slide and cover with a coverslip. Examine under low
power to be sure you have at least one cell.
2. View the Amoeba under high power. What structural features can you observe?
You will probably find the nucleus, food vacuoles, a variety of inclusions (many of
these are crystalline waste material), and maybe even a contractile vacuole.
3. A healthy Amoeba is quite active, extending pseudopodia in all directions and
creeping along the slide. This movement is accompanied by cytoplasmic streaming.
Look carefully at an extending pseudopod; notice that the inner core (endoplasm)
of the pseudopod is doing most of the streaming and is surrounded by a more rigid
shell of cytoplasm (ectoplasm). When the endoplasm reaches the advancing tip of
the pseudopod, what happens? Can you tell if the endoplasm is being "squirted"
forward into the pseudopod, or is it being "pulled" forward?
B. Paramecium
This organism is also a single cell with some of the same features as Amoeba. They
both move, eat and digest, excrete, remove water with a contractile vacuole, etc.,
but when you look at them, it seems hard to believe they are similar.
1. Add a small drop of culture water to a clean slide and mix in a drop of methyl
cellulose with a toothpick (this viscous substance will slow down paramecia
locomotion). Locate them with low power and then switch to high power (you will
have to continually move the slide to keep them in the field of view).
2. Try to locate the various internal structures, such as the nucleus (actually there is
a macronucleus and an associated micronucleus), food vacuoles, contractile
vacuoles (there are two, with radiating canals), and cytoplasmic inclusions. Watch
one of the contractile vacuoles for a few minutes. Do you see anything happening?
What is the frequency of the event? . Are the two vacuoles operating in synchrony?
Notice the large oral groove that tends to accumulate food particles for ingestion.
3. Do you see signs of cytoplasmic streaming? Does it contribute to the locomotion
of the animal? If not, what does cytoplasmic streaming do? The entire cell is
covered with cilia which beat in a coordinated sequence to propel the organism
along. Does the paramecium always swim in the same direction?

C. Human Cheek Epithelial Cells

Unlike Amoeba and Paramecium, the cells of multicellular organisms do not have
to perform all the functions of life - there is a "division of labor". As an example of
the "simpler" cells of multicellular organisms you will look at cells that line the
inside of your cheek. These cells are continually dying and being sloughed off, so
you shouldn't mind making this small contribution to science.
1. Using a clean toothpick, gently scrape the inside of your cheek and stir the cells
and a little saliva into a small drop of acetocarmine dye on the slide. Add a
coverslip and locate a group of flat and spread-out cells under low power.
2. Switch to a higher power for a better examination of the cells. What are their
shape? Do you see: the plasma membrane, a central vacuole, chloroplasts? You
should be able to see a nucleus and the nucleolus within it. What is the function of
the nucleolus?
3. Draw and label a cheek cell in the space below.

Abstract
To elucidate the role of cellulose microfibrils in the control of growth anisotropy, a link between their net
orientation, in vitro cell wall extensibility, and anisotropic cell expansion was studied during development
of the adaxial epidermis of onion (Allium cepa) bulb scales using polarization confocal microscopy, creep
tests, and light microscopy. During growth the net cellulose alignment across the whole thickness of the
outer epidermal wall changed from transverse through random to longitudinal and back to transverse
relative to the bulb axis. Cell wall extension in vitro was always higher transverse than parallel to the net
cellulose alignment. The direction of growth anisotropy was perpendicular to the net microfibril
orientation and changed during development from longitudinal to transverse to the bulb axis. The
correlation between the degree of growth anisotropy and the net cellulose alignment was poor. Thus the
net cellulose microfibril orientation across the whole thickness of the outer periclinal epidermis wall
defines the direction but not the degree of growth anisotropy. Strips isolated from the epidermis in the
directions perpendicular and transverse to a net cellulose orientation can be used as an extensiometric
model to prove a protein involvement in the control of growth anisotropy.

Related glassware
Main article: Laboratory flasks

 Flat-bottomed flask A flask with similar uses as the round-bottom flask, but the flat bottom allows it
to stand on a level surface.

 Florence flasks are similar flasks that have round bodies and either a round bottom or a flat
bottom so that one can stand the flask on a level surface. Florence flasks typically have one neck
which is longer and may be somewhat wider than the usual neck of a round bottom flask. The
 necks of traditional Florence flasks often don't have a ground glass joint like modern round bottom
flasks do. Round-bottom flasks are used more commonly by professional chemists than Florence
flasks.

 Retort a spherical vessel with a long downward-pointing neck, specially used for distillation or dry
distillation of substances.

 Schlenk flask - a round-bottom flask with a built-in plug valve or stopcock.

[hide]

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cylinder · Pipette · Petri dish · Pycnometer · Separatory funnel · Soxhlet extractor · Watch glass

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are
ksBüchner · Erlenmeyer · Fleaker · Florence · Retort · Round-bottom · Schlenk · Volumetric

Tub
esBoiling · NMR · Test · Thiele · Thistle

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stirrer · Meker-Fisher burner · Microscope · Microtiter plate · Picotiter plate · Plate reader · Retort stand · Safety shower ·Spectrophotome

bar · Stirring rod · Scoopula · Thermometer · Vortex mixer · Wash bottle

oratory glassware
From Wikipedia, the free encyclopedia

It has been suggested that Ground glass joint be merged into this
article or section. (Discuss)
Brown glass jars with some clear lab glassware in the background

Three beakers, a conical flask, a graduated cylinder and a volumetric flask

Laboratory glassware refers to a variety of equipment, traditionally made of glass, used forscientific
experiments and other work in science, especially in chemistry and biologylaboratories. Some of the
equipment is now made of plastic for cost, ruggedness, and convenience reasons, but glass is still used for
some applications because it is relativelyinert, transparent, more heat-resistant than some plastics up to a
point, and relatively easy to customize. Borosilicate glasses—formerly called Pyrex—are often used
because they are less subject to thermal stress and are common for reagent bottles. For some
applicationsquartz glass is used for its ability to withstand high temperatures or its transparency in certain
parts of the electromagnetic spectrum. In other applications, especially some storagebottles, darkened
brown or amber (actinic) glass is used to keep out much of the UV and IR radiation so that the effect of light
on the contents is minimized. Special-purpose materials are also used; for example, hydrofluoric acid is
stored and used in polyethylene containers because it reacts with glass.[1] For pressurized reaction, heavy-
wall glass is used forpressure reactor.

Contents
 [hide]

• 1 Applications

• 2 Production

• 3 Service temperatures

• 4 Lubrication and sealing

• 5 Safety when using vacuums and

Keck clips

• 6 Gentle & even heating - baths &

alternatives

• 7 Glassware Joints

○ 7.1 Ground glass joints

 7.1.1 Conically

tapered joints

 7.1.2 Ball-and-

socket joints

○ 7.2 O-ring joints

○ 7.3 Threaded connections

○ 7.4 Glass-to-metal

transition joints

○ 7.5 Hose connections

• 8 Glassware Valves

○ 8.1 Stopcock valve

○ 8.2 Threaded plug valve

• 9 Fritted glass

• 10 Cleaning laboratory glassware

• 11 Gallery

• 12 Notes

[edit]Applications

There are many different kinds of laboratory glassware items, the majority are covered in separate articles
of their own; see the list further below. Such glassware is used for a wide variety of functions which include
volumetric measuring, holding or storing chemicals or samples, mixing or preparing solutions or other
mixtures, containing lab processes like chemical reactions, heating, cooling, distillation, separations
including chromatography, synthesis, growing biological organisms, spectrophotometry, and containing a
full or partial vacuum, and pressure, like pressure reactor. When in use, laboratory glassware is often held
in place with clamps made for that purpose, which are likewise attached and held in place by stands or
racks. This article covers aspects of laboratory glassware which may be common to several kinds of
glassware and may briefly describe a few glassware items not covered in other articles.

[edit]Production

Most laboratory glassware is now mass-produced, but many large laboratories employ a glass blower to
construct specialized pieces. This construction forms a specialized field of glassblowing requiring precise
control of shape and dimension. In addition to repairing expensive or difficult-to-replace glassware, scientific
glassblowing commonly involves fusing together various glass parts—such as glass joints and tubing,
stopcocks, transition pieces, and/or other glassware or parts of them to form items of glassware, such
as vacuum manifolds, special reactionflasks, etc.

Various types of joints and stopcocks are available separately and come fused with a length of glass tubing,
which a glassblower may use to fuse to another piece of glassware.

[edit]Service temperatures

Borosilicate glass, which makes up the majority of lab glass, may fracture if rapidly heated or cooled
through a 150 °C (302 °F) temperature gradient. This is particularly true of large volume flasks, that can
take hours to safely warm up. Gentle thermal cycling should be used when working with volumes more than
hundreds of mls to two liters. Whenever working with borosilicate glass, it is advisable to avoid sharp
transitions between temperatures when the heating and cooling elements have a high thermal inertia.
Glassware can be wrapped with tinfoil or insulated with wool to smooth out temperature gradients.

500 °C (932 °F) is the maximum service temperature for borosilicate glass as, at 510 °C (950 °F), thermal
strain begins to appear in the structures. Operation at this temperature should be avoided and only
intermittent. Bear in mind that glassware under vacuum will also have around one atmosphere of pressure
on its surface before heating and so will be more likely to fracture as temperature transitions increase.
Vacuum operation should be used if the atmospheric temperatures required are above a few hundred
degrees Celsius, as this often has a dramatic effect on boiling points; significantly lowering them.

Borosilicate anneals at 560 °C (1,040 °F), this removes built in stain in the glass.

At 820 °C (1,510 °F), borosilicate glass softens and is likely to deform. And at 1,215 °C (2,219 °F) it
becomes workable.

Quartz glass is far more resilient to thermal shock and can be operated continuously at 1,000 °C (1,830 °F).
Thermal strain appears at 1,120 °C (2,050 °F), annealing occurs at 1,215 °C (2,219 °F) and it becomes
workable at 1,685 °C (3,065 °F).

It is common for students and those new to working with glassware to set hotplates to a high value initially
to rapidly warm a solution or solid. This is not only bad practice, as it can scorch the contents, it will almost
universally burst large flasks, and this is one of the reasons why large flasks are often heated in water, oil,
sand and steam baths or using a mantle that surrounds most, or all, of the flask.

[edit]Lubrication and sealing

A thin layer of grease is usually applied to the ground-glass surfaces to be connected, and the inner joint is
inserted into the outer joint such that the ground-glass surfaces of each are next to each other to make the
connection. The use of grease helps to provide a good seal and prevents the joint from seizing, allowing the
parts to be disassembled easily.[2]

Grease can be washed out of tapers by the flow of solvents past them. Reagents may react with the grease
or it may leak from the tapers at higher temperatures. The latter is prone to occurring when the system is
under vacuum. Grease leaking from tapers can, of coarse, contaminate an operation either passively or by
actively reacting with something passing by. For these reasons, it is advisable to apply a light ring of grease
at the fat end of the taper and not its tip, to keep the material away from insides of the glassware. If the
grease smears over the entire taper surface on mating, too much is being used. Using greases specifically
designed for this purpose is also a good idea, as these are often better at sealing under vacuum, thicker
and so less likely to flow out of the taper, become fluidic at higher temperatures than Vaseline (a common
substitute) and are more chemically inert than other substitutes.

Grease allows chemists to easily see when a taper is leaking, as bubbles can usually be seen flowing
through the taper.

When contamination is a serious concern, PTFE (Teflon) sleeves and PTFE sealing rings can be used in
between joints to fit them together instead of grease.[3] PTFE tape can also be used, but requires a little care
when winding onto the joint to ensure a good seal is produced.

Keck clips and other clamping methods can be used to hold glassware together.

[edit]Safety when using vacuums and Keck clips

An absolute vacuum produces a pressure of one atmosphere, approximately 14 psi, over the surface of the
glass. The energy contained within an implosion is defined by the pressure difference and the volume
evacuated. As most vacuum chemistry occurs with at least 90% of the atmosphere removed, the pressure
difference is often negligible between laboratories; a typical diphragm pump, aspirator or rotary vane pump
will remove this level of atmosphere. However, flaks volumes can change by orders of magnitude between
experiments. Whenever working with liter sized or larger flasks, chemists should consider using a safety
screen or the sash of a flow hood to protect them from shards of glass, should an implosion occur.
Glassware can also be wrapped with spirals of tape to catch shards, or wrapped with webbed mesh more
commonly seen on scuba cylinders.

Glass under vacuum becomes more sensitive to chips and scratches in its surface, as these form strain
accumulation points, so older glass is best avoided if possible. Impacts to the glass and thermally induced
strain are also concerns under vacuum.
Round bottom flasks more effectively spread the strain across their surfaces, and are therefore safer when
working under vacuum. For the majority of work below a liter, the difference in flask form has little influence
on its safety under vacuum; Erlenmeyer flasks can be used.

When connecting glassware, it is often tempting to use Keck clips on every joint, but this can be dangerous
if the system is sealed or the exhaust is in anyway restricted; e.g. by wash flasks or drying media. Many
reactions and forms of operation can produce sudden, unexpected surges of pressure inside the glass. If
the system is sealed or restricted, this can blow the glass apart. It is safer to only clip the joints that need
holding together to stop them falling apart and to purposefully leave one or more unclipped; preferably
those that are connected to lightweight, small objects like stoppers, thermometers or wash heads, that are
pointing vertically upwards and not connected to other items of glassware. By doing so, any significant
surge of pressure will cause these specifically chosen tapers to open and vent. This may seem
counterintuitive, but it is safer and easier to deal with a controlled escape as opposed to the entire volume
being uncontrollably released in an explosion.

[edit]Gentle & even heating - baths & alternatives

This is a prerequisite for a lot of laboratory work as it protects the work itself and decreases the possibility of
thermal strain fracturing the glass; see service temperatures for more information on this.

A common method is to fill a bowl surrounding the flask with water, oil, sand or steam, or to use a wrap
around heating mantle.

However, baths can be extremely dangerous if they spill, overheat or ignite, they have a high thermal inertia
(and so take a long time to cool down) and mantles can be very expensive and are designed for specific
flask volumes. There are two alternative methods that can be used instead, where appropriate.

When a heat sources minimum temperature is high, the glassware can be suspended slightly above the
surface of the plate. This will not only reduce the ultimate temperature on the glass, it will slow down the
rate of heat exchange and encourage more even heating; as there is no longer direct contact via a few
points with the plate. Doing so works well for low boiling point operations.

If the glassware must be run at higher temperatures, a teepee setup can be used; so named as it looks a
little like a tipi. This is when the glassware is suspended above the plate, but the flask is surrounded by a
skirt of tinfoil. The skirt should start at the neck of the flask and drape down to the surface of the plate, not
touching the sides of the flask. Having the base of the skirt cover the majority of the plates surface will effect
better heat transfer. The flask will now be warmed indirectly by the hot air collecting under the skirt but,
unlike simply suspending the glassware, it can now reach hundreds of degrees Celsius and is better
protected from drafts.

Both these methods are useful as they are either cheaper or free, effective, safe and feature low thermal
inertia transfer methods, meaning the chemist does not have to wait for a bath to cool down after use.
Baths are most useful when the heat source has little or no control over it. With the advent of variable
temperature hotplates and wrap around mantles, their necessity has somewhat declined. The same can be
said for many round bottom flask operations, which require the use of a bath.

[edit]Glassware Joints
[edit]Ground glass joints
Main article: Ground glass joint

In a lab experiment or process—such as a distillation or a reflux—ground glass joints make it possible to

rapidly assemble the set-up from component glassware items in a leak-tight but non-permanent way. Using
old technology, this was often done with rubber (or possibly cork) stoppers inserted between the component
glassware items. Holes could be made in such stoppers to insert glass tubes or the ends of some glass
items. However, rubber (and of course cork) are not as chemically inert or heat-resistant as glass and
degrade with age. In order to connect the hollow inner spaces of the glassware components, these types of
joints are hollow on the inside and open at the ends, except for stoppers.

Two general types of ground glass joints are fairly commonly used: joints that are slightly conically-tapered
and ball and socket joints (sometimes called spherical joints).

Ground glassware should be disassembled as soon as it is safe to do so after a reaction, as this will help
avoid the tapers seizing. Tapers will seize either from thermal activity or something from the reaction
penetrating the taper and thickening upon cooling or exposure to the atmosphere. High concentrations of
certain reactants can chemically seize a taper such that is essentially impossible to open. Exposure time
plays a role in this occurring, and so should be minimized. PTFE sealing methods are also of use for such
applications as they produce an intermediate layer between the glass that is highly inert and solid, meaning
it can not be displaced and the glass can never come in contact with its mating taper.

[edit]Conically tapered joints

Conically tapered ground glass joints consist of a male and a female half[2] which are manufactured to a
standard 1:10 taper. Apart from stoppers, most conically tapered joints are hollow to allow liquids or gases
to flow through. An example of the use of conically-tapered joints is to join a round bottom flask, Liebig
condenser, and oil bubbler together to allow a reaction mixture to be refluxed.

Conically tapered ground glass joints. Inner (male) joint shown on the left and outer (female) joint shown on the right.
Ground glass surfaces are shown with gray shading. By putting them together in the direction of the arrows, they can be
joined, usually with some grease applied to the ground glass surfaces.
[edit]Ball-and-socket joints

Here, the inner joint is a ball and the outer joint is a socket, both having holes leading to the interior of their
respective tube ends to which they are fused. Ball and socket joints are used where some degree of free-
play is necessary, such as when joining a cold trap to a gas manifold for a Schlenk line.[2]

Ground glass ball (left) and socket (right) joints. The ground glass surfaces are shown with gray shading. By putting
them together in the direction of the arrows, they can be joined, with some grease applied to the ground glass surfaces.

For either standard taper joints or ball-and-socket joints, inner and outer joints with the same numbers are
made to fit together. When the joint sizes are different, ground glass adapters may be available (or made) to
place in between to connect them. Special clips or pinch clamps, known as Keck clips, may be placed
around the union of the joints to help keep them together.

Grease is used to lubricate glass stopcocks and joints. Some laboratories fill them into syringes for easy application.
Two typical examples: Left - Krytox, a fluoroether-based grease; Right - a silicone-based high vacuum grease by Dow
Corning.

[edit]O-ring joints
There are also glass joints available sometimes which use an O-ring between them to form a leak-tight seal.
[2]
Such joints are more symmetrical in theory with a tubular joint on each side having a widened tip with a
concentric circular groove into which an elastomer O-ring can be inserted between the two joints. O-ring
joints are sized based on the inner diameter in mm of the joint. Since they can come apart rather easily, a
clip or pinch clamp is needed to hold them together. The elastomer of the O-ring is more limited in
high temperature resistance than other types of glass joints using high temperature grease.
Glass O-ring joints with elastomer O-ring in between. By putting them together in the direction of the arrows with an
appropriately-sized O-ring placed in between in circular grooves on each joint (not shown on the joint on the left side for
simplicity), they can be joined.

[edit]Threaded connections
Round slightly spiral threaded connections are possible on tubular ends of glass items. Such glass
threading can face the inside or the outside. In use, glass threading is screwed into or onto non-glass
threaded material such as plastic. Glass vials typically have outer threaded glass openings onto which caps
can be screwed on. Bottles and jars in which chemicals are sold, transported, and stored usually have
threaded openings facing the outside and matching non-glass caps or lids.

[edit]Glass-to-metal transition joints

Occasionally, it may be desired to fuse a glassware item to a metal item with a tubular pathway between
them. This requires the use of a glass-to-metal transition joint. Most glass used in laboratory glassware
does not have the same coefficient of thermal expansion as metal, so fusing the usual type of glass with
metal is likely to result in cracking of the glass. These special transition joints have several short sections of
special types of glass fused together between the metal and the usual type of glass, each having more
gradual changes in thermal expansion coefficients.

[edit]Hose connections
Laboratory glassware, such as Buchner flasks and Liebig condensers, may have tubular glass tips serving
as hose connectors with several ridged hose barbs around the diameter near the tip. This is so that the tips
can have the end of a rubber or plastic tube mounted over them to connect the glassware to another
system such as a vacuum, water supply, or drain. A special clip may be placed over the end of the flexible
tube surrounding the connector tip to prevent the hose from slipping off the connector.

A number of brands, including Quickfit, have begun using threaded connections for hose barbs. This allows
the barb to be unscrewed from the glassware, the hose pushed on and the setup screwed back together.
This helps avoid accidentally breaking the glass and potentially doing serious harm to the chemist, as will
sometimes occur when pushing the hoses directly onto the glass.
[edit]Glassware Valves

A very common straight bore glass stopcock attached with a plastic plug retainer. This stopcock is in the side arm of a
Schlenk flask.

Describing glassware can be complicated since manufactures provide conflicting names for glassware. For
example ChemGlass calls a glass stopcock what Kontes calls a glass plug. Despite this it is clear there are
two main types of valves used in laboratory glassware, thestopcock valve and the threaded plug valve.
These and other terms used below are defined in detail since they are bound to conflict with different
sources.

[edit]Stopcock valve
Stopcocks are often parts of laboratory glassware such as burettes, separatory funnels, Schlenk flasks, and
columns used for column chromatography. The stopcock is a smooth tampered plug or rotor with a handle,
which fits into a corresponding ground glass female joint. The stationary female joint is designed such that it
joins two or more pieces of glass tubing. The stopcock has holes bored through it which allow the tubes
attached to the female joint to be connected or separated with partial turns of the stopcock. Most stopcocks
are solid pieces with linear bores although some are hollow with holes to simple holes that can line up the
joints tubing. The stopcock is held together with the female joint with a metal spring, plastic plug retainer, a
washer and nut system, or in some cases vacuum. Stopcocks plugs are generally made out of ground glass
or an inert plastic like PTFE. The ground glass stopcocks are greased to create an airtight seal and prevent
the glass from fusing. The plastic stopcocks are at most lightly oiled.

Stopcocks are generally available individually with some length of glass tubing at the ports so that they can
be joined by a glass blower into custom apparatus at the point of use. This is especially common for the
large glass manifolds used in high vacuum lines.

More examples are featured in the gallery. This is a small sampling of stopcock valves; many additional
variations exist in both plug boring and joint assembly.

[edit]Threaded plug valve

A standard solid threaded plug valvewith a double o-ring upper seal and PTFE to glass seal at it base.

Threaded plug valves are used significantly in air-sensitive chemistry as well as when a vessel must be
closed completely as in the case of Schlenk bombs. The construction of a threaded plug valve involves a
plug with a threaded cap which are made so that they fit with the threading on a corresponding pieces of
female glass. Screwing the plug in part way first engages one or more o-rings, made of rubber or plastic,
near the plugs base which seals the female joint off from the outer atmosphere. Screwing the plug valve all
the way in engages the plugs tip with a beveled constriction in the glass which provides a second seal. This
seal separates the region beyond the bevel and the o-rings already mentioned.

With solid plugs a tube or area exists above and below the bevel and turning the plug controls access. In a
number of cases its convent to fully remove a plug which can give access to the region beyond the bevel.
Plugs are generally made of an inert plastic such as PTFE with and are attached to a threaded sleeve in
such a way that the sleeve can been turned without spinning the plug. The contact with the bevel is made
by an o-ring fitted to the tip of the plug or by the plug itself. There are a few examples where the plug in
made of glass. In the case of glass plugs the joint contact is always a rubber o-ring but are still prone to
shattering.
A thread T-bore plug valve used as a side arm on a Schlenk flask.

Not all plugs are solid. Some plugs are bored with a T-junction. In these systems the plug extends beyond
the threaded sleeve and is designed to form an airtight fitting with glass tubing or hosing. The shaft of the
plug is bored from beyond the threaded sleeve to a T-junction just before the bevel plug contact. When the
plug is fully sealed region beyond the bevel is separated from the plug shaft as well as the bore which leads
out of its shaft. When the plug bevel contact is released the two regions are exposed to each other. These
valves have also be used as a grease free alternative to straight bored stopcocks common to Schlenk
flasks. The high symmetry and concise design of these valves has also made them popular for
capping NMR tubes. Such NMR tubes can be heated without the loss of solvent thanks to the valve's gas
tight seal. NMR tubes with T-bore plugs are widely known as J. Young NMR tubes named after the brand
name of valves most commonly used for this purpose. Images of J. Young NMR tubes and a J. Young NMR
tube adapter are in the gallery.

[edit]Fritted glass

A Büchner funnel with a sintered glass disc

Fritted glass is finely porous glass through which gas or liquid may pass. It is made by sintering together
glass particles into a solid but porous body.[4]This porous glass body can be called a frit. Applications in
laboratory glassware include use in fritted glass filter items, scrubbers, or spargers. Other laboratory
applications of fritted glass include packing in chromatography columns and resin beds for special chemical
synthesis.

In a fritted glass filter, a disc or pane of fritted glass is used to filter out solid particles, precipitate, or residue
from a fluid, similar to a piece of filter paper. The fluid can go through the pores in the fritted glass, but the
frit will often stop a solid from going through. A fritted filter is often part of a glassware item, so fritted glass
funnels and fritted glass crucibles are available.[5]

Gas-washing bottle

Laboratory scale spargers (also known as gas diffusing stones or diffusors) as well as scrubbers, and gas-
washing bottles (orDrechsel bottles [6]) are similar glassware items which may use a fritted glass piece fused
to the tip of a gas-inlet tube. This fritted glass tip is placed inside the vessel with liquid inside during use
such that the fritted tip is submerged in the liquid. To maximize surface area contact of the gas to the liquid,
a gas stream is slowly blown into the vessel through the fritted glass tip so that it breaks up the gas into
many tiny bubbles. The purpose of sparging is to saturate the enclosed liquid with the gas, often to displace
another gaseous component. The purpose of a scrubber or gas-washing bottle is to scrub the gas such that
the liquid absorbs one (or more) of the gaseous components to remove it from the gas stream, effectively
purifying the gas stream.

As frits are made up of particles of glass that are bonded together by small contact areas, it is wise to avoid
using them in strongly alkaline conditions, as these can dissolve the glass to some extent. This is not
normally a problem, as the amount dissolved is usually minute, but the equally minute bonds in a frit can be
rotted away, causing the frit to fall apart over time. As such, consideration should be given to using frits in
such solutions and they should be rapidly and thoroughly rinsed when cleaning the glass with bases like
KOH.
[edit]Cleaning laboratory glassware

There are many different methods of cleaning laboratory glassware. Most of the time, these methods [7]
[8]
are tried in this order:

 The glassware is soaked in a detergent solution to remove grease and loosen most contamination

 Gross contamination and large particles are removed mechanically, by scrubbing with a brush or
scouring pad.

 Alternatively, the first two steps may be combined by sonicating the glassware in a hot detergent
solution

 Solvents known to dissolve the contamination are used to rinse the glassware and remove the last
traces

 Acetone is often used for a final rinse of sensitive or urgently needed glassware as the solvent
is miscible with water and forms a lowboiling point azeotrope with it, encouraging the remaining
aqueous phase to leave more rapidly and thoroughly; this is particularly important if the following work
is moisture sensitive.

 Glassware is often dried by suspending it upside down to drip dry on racks; these can include a hot air
fan to blow the internals dry. Another alternative is to place the glassware under vacuum, lower the
boiling points of the remaining volatiles.

If the glassware are still dirty, more drastic methods may be needed. This includes soaking the piece in a
saturated solution of sodium orpotassium hydroxide in an alcohol ("base bath"),[8] followed by a dilute
solution of hydrochloric acid ("acid bath") to neutralize the excess base. Sodium hydroxide cleans glass by
dissolving a tiny layer of silica, to give soluble silicates. Care should be taken using strongly alkaline
solutions to clean fritted glassware, as this will degrade the frit over time.

More aggressive methods involving aqua regia (for removing metals from frits), piranha
solution and chromic acid (for removing organics), andhydrofluoric acid baths are generally considered
unsafe for routine use because of possible explosions and the corrosive/toxic materials involved.[8]

[edit]Gallery

A single hole hollow glass

A double oblique bore glass
stopcock held in place by
A straight bore plastic three-way stopcock.
A T-bore glass stopcockin a vacuum.
stopcock sans female joint.
three way assembly. Two of the
Note its washer and nut system
for attaching to its female joint. outlets end in plain hose adapters

while the third ends in a male

14/20 ground glass joint. This

stopcock is attached with an

easily removed metal spring.

A J. Young NMR tube from

A J. Young NMR above looking down the hole that A Taper Joint Stopper with
tubeattached to an adapter with leads to the T-bore. PTFE Sealing Ring. Optical
a female 24/40 joint already transparency of the narrow
greased. Note the hole sealing ring pressured by
resulting from the T-bore in the glass joint (right).
side of the PTFE plug.

Materials and methods

7. First give a list of all the equipment used in the experiment. Give the
size of beakers/measuring cylinders, etc, used, give the names of any
chemicals that are used in the experiment.

8. You can use a diagram (picture) to show the experimental set up if you
find it necessary.

9. Now you should describe the method. It should be written in past tense
(i.e. not written as a guide on how to carry out the experiment again, but
rather, how you did it). The steps in the experiment are either self-evident
or explained.

10.In this part you should explain the different variables. Write how the
independent variable was varied. Using the yeast example, the
independent variable can be varied by placing the fermentation tubes in
hot water baths of different temperatures.

11.Write how changes of the dependent variable were monitored. You

should write how you got your results, e.g. by reading from the scale on
the fermentation tube to see how much CO2 that has been produced.
 12.Write how the controlled variables were controlled. Using the yeast
example, you write that you made sure that the amount of yeast used in
each fermentation tube was the same (because you used a scale), that
you used a watch to make sure that the time that the tubes were allowed
to ferment was the same for all tubes.

13.Write how you made sure that the sufficient relevant data was
recorded. Describe the method for data collection, i.e. if you had several
trials, if you used controls, methods of measurements, if your calculations
are correct, etc.

Results
Data collection

14.Record all your raw data in tables. The tables should be numbered and
have captions in which you briefly describe the contents of the tables and
how you recorded the results. Titles, units and the uncertainty should be
given in the headings of the tables.
15.Underneath the table you can briefly describe the results. You can
describe the main trends and account for any anomalous result. You don’t
have to discuss the significance of the results to the aim of the
investigation.

Data Processing and presentation

16.The data should be processed (calculated) correctly and presented in

tables (as above) and graphs. If you use graphs, they should have a
caption in which you describe the contents of the graph. The axes of the
graphs have to be labelled with units and the points have to be plotted
correctly. Make sure that you use the correct type of graphs. If both
variables are continuous, use a point graph.
17.For HL: Error analysis should be carried out if possible (calculate the
percentage uncertainty, etc).

Conclusion
18.In the conclusion you should discuss the results you obtained in relation
with your hypothesis. Write a conclusion based on an interpretation of the
gathered results.
19.Compare your results with literature values if possible.

Evaluation
 20.In the evaluation you should evaluate the method used. Write about the
main weakness of the method used and the weakness in the method of
manipulation of data.
21.Write about the source of error, but don’t include personal mistakes.
22.Suggest real improvements (that can be carried out in the school lab) to
the investigation.
23.Discuss further investigations that are of interest and can be carried out
and new questions that could be posed.

Written by: IB Genius

Back

A fleaker is a type of container for liquids used in the laboratory. It can be described as a cross
between the Griffin beaker and the Erlenmeyer flask.

Like a beaker, the bottom is flat, with the sides meeting the bottom at a 90 degree angle. The sides
are vertical for most of the height; near the top, the sides curve in to form a neck with a widely flared
rim. The wide rim makes it easier to pour from or filter into; the narrow neck reduces loss of the
contents due to splashing and serves as a grip for handling and pouring. Fleakers have a plastic lid
with a built in rubber stopper. When on the fleaker, the lid covers the narrow neck. Fleakers work as
well as other glassware for liquids and solutions, but are inappropriate for slurries, precipitates, and
recrystallizations (since the narrow neck makes it difficult to remove solids completely from a fleaker).

The fleaker was invented by Roy Eddleman, founder of Spectrum Medical Industries (now Spectrum
Laboratories).

Beaker (glassware)
From Wikipedia, the free encyclopedia

Beaker
 Beakers of several sizes

Uses Liquid volume containment

and measurement

Related items Fleaker

A beaker is a simple container for stirring, mixing and heating liquids commonly used in many laboratories.
Beakers are generally cylindrical in shape, with a flat bottom and a lip for pouring.[1]Many also have a small
spout to aid pouring as shown in the picture. Beakers are available in a wide range of sizes, from
one millilitre up to several litres.

Contents

[hide]

• 1 Structur

• 2 Materials

• 3 Shape

• 4 See also

• 5 Referenc

es

• 6 Further

reading

[edit]Structure

Standard or "Low-form" beakers typically have a height about 1.4 times the diameter.[2] The common low
form with a spout has been called the Griffin form.[3] (A in the image) These are the most universal character
and are used for various purposes - from preparing solutions and decanting supernatant fluids to simple
reactions.

Beakers

"Tall form" beakers have a height about twice the diameter.[2] These are sometimes called Berzelius
beakers; (B in the image). They are mostly used for titration.

Flat beakers (C in the image) are often called crystallizers, because most are used to perform
crystallization, but often it is also used as a vessel for use in hot-bath heating. These beakers usually do not
have a flat scale.

A beaker is distinguished from a flask by having sides which are straight rather than sloping. The exception
to this definition is a slightly conical sided beaker called a Phillips beaker.

[edit]Materials

Beakers are commonly made of glass (today usually borosilicate glass[2]), but can also be in metal (such
as stainless steel or aluminium) or certain plastics (notably polythene, polypropylene, PTFE). A common
use for polypropylene beakers is gamma spectral analysis of liquid and solid samples.

[edit]Shape

Beakers are often graduated, that is, marked on the side with lines indicating the volume contained. For
instance, a 250 mL beaker might be marked with lines to indicate 50, 100, 150, 200, and 250 mL of volume.
These marks are not intended for obtaining a precise measurement of volume (a graduated cylinder or
a volumetric flask would be a more appropriate instrument for such a task), but rather an estimation.

The presence of a lip means that the beaker cannot have a lid. However, when in use, beakers may be
covered by a watch glass to prevent contamination or loss of the contents, but allowing venting via the
spout.
[edit]See also
 Fleaker

 Stirring rod
[edit]References

Laboratory equipment refers to the various tools and equipment used by scientists working in
a laboratory. These include tools such asBunsen burners, and microscopes as well as specialty
equipment such as operant conditioning chambers, spectrophotometers andcalorimeters. Another
important type of laboratory equipment is Laboratory glassware.

Laboratory equipment is generally used to either perform an experiment or to take measurements and
gather data. Larger or more sophisticated equipment is generally called a scientific instrument.

[edit]See also

IB Internal Assessment Rubrics and guidelines

 IB Biology HL

The Diploma Programme hexagon

The course is presented as six academic areas enclosing a central core. It encourages the concurrent study of a br
academic areas. Students study: two modern languages (or a modern language and a classical language); a huma
science subject; an experimental science; mathematics; one of the creative arts. It is this comprehensive range of
the Diploma Programme a demanding course of study designed to prepare students effectively for university entra
academic areas students have flexibility in making their choices, which means they can choose subjects that partic
and that they may wish to study further at university.

HL group 4 curriculum model

HL Total teaching hours 240

Theory 180
Core 80

Additional higher level (AHL) 55

Options 45

Practical work 60
Investigations 50

Group 4 project 10

The objectives for all group 4 subjects reflect those parts of the aims that will be assessed. Wherever appropriate,
draw upon environmental and technological contexts and identify the social, moral and economic effects of science
It is the intention of all the Diploma Programme experimental science courses that students achieve the following o
1. Demonstrate an understanding of:
a. scientific facts and concepts
b. scientific methods and techniques
c. scientific terminology
d. methods of presenting scientific information.
2. Apply and use:
a. scientific facts and concepts
b. scientific methods and techniques
c. scientific terminology to communicate effectively
d. appropriate methods to present scientific information.
3. Construct, analyse and evaluate:
a. hypotheses, research questions and predictions
b. scientific methods and techniques
c. scientific explanations.
4. Demonstrate the personal skills of cooperation, perseverance and responsibility appropriate for effect
investigation and problem solving.
5. Demonstrate the manipulative skills necessary to carry out scientific investigations with precision and

The external assessment consists of three written papers.

Paper 1
Paper 1 is made up of multiple-choice questions that test knowledge of the core only for students at SL and the cor
for students at HL. The questions are designed to be short, one- or two-stage problems that address objectives 1 a
“Objectives” section). No marks are deducted for incorrect responses. Calculators are not permitted, but students
out simple calculations.

Paper 2
Paper 2 tests knowledge of the core only for students at SL and the core and AHL material for students at HL. The q
objectives 1, 2 and 3 and the paper is divided into two sections.
In section A, there is a data-based question that requires students to analyse a given set of data. The remainder of
of short-answer questions.
In section B, students at SL are required to answer one question from a choice of three, and students at HL are req
questions from a choice of four. These extended-response questions may involve writing a number of paragraphs,
problem, or carrying out a substantial piece of analysis or evaluation. A calculator is required for this paper.

Paper 3
Paper 3 tests knowledge of the options and addresses objectives 1, 2 and 3. Students at SL are required to answer
questions in each of the two options studied. Students at HL are required to answer several short-answer questions
response question in each of the two options studied. A calculator is required for this paper.

Internal Assessment Criteria:

Criteria and aspects
There are five assessment criteria that are used to assess the work of both SL and HL students.
• Design—D
• Data collection and processing—DCP
• Conclusion and evaluation—CE
• Manipulative skills—MS
• Personal skills—PS
The first three criteria—design (D), data collection and processing (DCP) and conclusion and evaluation (CE)—are e
Manipulative skills (MS) is assessed summatively over the whole course and the assessment should be based on a
manipulative skills.
Personal skills (PS) is assessed once only and this will be during the group 4 project.
Each of the assessment criteria can be separated into three aspects as shown in the following sections. Descriptio
indicate what is expected in order to meet the requirements of a given aspect completely (c) and partially (p). A
given for circumstances in which the requirements are not satisfied, not at all (n).
A “complete” is awarded 2 marks, a “partial” 1 mark and a “not at all” 0 marks.
The maximum mark for each criterion is 6 (representing three “completes”).
D ×2=
12

DC ×2=
P 12

CE ×2=
12

MS ×1=
6

PS ×1=
6

This makes a total mark out of 48.

The marks for each of the criteria are added together to determine the final mark out of 48 for the IA component. T
IBCA to give a total out of 24%.
General regulations and procedures relating to IA can be found in the Vade Mecum for the year in which the IA is b
Design

Levels/ma Aspect 1 Aspect 2 Aspect 3

rks
Defining the problem and Controlling variables Developing a me
selecting variables collection of dat

Complete/ Formulates a focused problem/research Designs a method for the effective Develops a method
2 question and identifies the relevant control of the variables. collection of suffici
variables.

Partial/1 Formulates a problem/research Designs a method that makes some Develops a method
question that is attempt to control the variables. collection of insuffi
incomplete or identifies only some
relevant variables.

Not at all/0 Does not identify a problem/research Designs a method that does not control Develops a method
questionand does not identify any the variables. for any relevant da
relevant variables.

Data collection and processing

Levels/ma Aspect 1 Aspect 2 Aspect 3

rks
Recording raw data Processing raw data Presenting proce

Complete/ Records appropriate quantitative and Processes the quantitative raw data Presents processed
2 associated qualitative raw data, correctly. and, where relevan
including units and uncertainties where and uncertainties.
relevant.

Partial/1 Records appropriate quantitative and Processes quantitative raw data, but Presents processed
associated qualitative raw data, but with some mistakes and/or omissions. but with some mist
with some mistakes or omissions. omissions.

Not at all/0 Does not record any appropriate No processing of quantitative raw data Presents processed
quantitative raw data or raw data is is carried out or major mistakes are inappropriately ori
incomprehensible. made in processing.

Conclusion and evaluation

Levels/ma Aspect 1 Aspect 2 Aspect 3

rks
Concluding Evaluating procedure(s) Improving the in

Complete/ States a conclusion, with justification, Evaluates weaknesses and limitations. Suggests realistic i
2 based on a reasonable interpretation of respect of identifie
the data. limitations.

Partial/1 States a conclusion based on a Identifies some weaknesses and Suggests only supe
reasonable interpretation of the data. limitations, but the evaluation is weak improvements.
or missing.

Not at all/0 States no conclusion or the conclusion Identifies irrelevant weaknesses and Suggests unrealisti
is based on an unreasonable limitations.
interpretation of the data.

Manipulative skills (assessed summatively)

This criterion addresses objective 5.
 Levels/ma Aspect 1 Aspect 2 Aspect 3
rks
Following instructions* Carrying out techniques Working safely

Complete/ Follows instructions accurately, Competent and methodical in the use Pays attention to s
2 adapting to new circumstances of a range of techniques and
(seeking assistance when required). equipment.

Partial/1 Follows instructions but requires Usually competent and methodical in Usually pays attent
assistance. the use of a range of techniques and
equipment.

Not at all/0 Rarely follows instructions or requires Rarely competent and methodical in Rarely pays attenti
constant supervision. the use of a range of techniques and
equipment.

*Instructions may be in a variety of forms: oral, written worksheets, diagrams, photographs, videos, flow charts, au
computer programs, and so on, and need not originate from the teacher.

Design
Aspect 1: defining the problem and selecting variables
It is essential that teachers give an open-ended problem to investigate, where there are several independent varia
student could choose one that provides a suitable basis for the investigation. This should ensure that a range of pla
formulated by students and that there is sufficient scope to identify both independent and controlled variables.
Although the general aim of the investigation may be given by the teacher, students must identify a focused probl
research question. Commonly, students will do this by modifying the general aim provided and indicating the varia
investigation.
The teacher may suggest the general research question only. Asking students to investigate some property of a pl
variables are given, would be an acceptable teacher prompt. This could be focused by the student as follows: “Doe
chloroplasts in Elodea leaf cells vary with light intensity?”
Alternatively, the teacher may suggest the general research question and specify the dependent variable. An exam
teacher prompt would be to ask the student to investigate the effect of a factor that influences enzyme activity. Th
focused by the student as follows: “Does ethanol concentration affect the activity of bovine catalase?” It is not suff
merely to restate the research question provided by the teacher.
Variables are factors that can be measured and/or controlled. Independent variables are those that are manipulate
this manipulation leads to the measurement of the dependent variable. A controlled variable is one that should be
not to obscure the effects of the independent variable on the dependent variable.
The variables need to be explicitly identified by the student as the dependent (measured), independent (manipulat
variables (constants). Relevant variables are those that can reasonably be expected to affect the outcome. For exa
investigation “How does the speed of movement of chloroplasts in Elodeacells vary with light intensity?”, the stude
that the independent variable is the light intensity and the dependent variable is the speed of movement. Relevan
would include temperature, preparation of Elodea cells, sample size and light quality (wavelength).
Students should not be:
• given a focused research question
• told the outcome of the investigation
• told which independent variable to select
• told which variables to hold constant.

Aspect 2: controlling variables

“Control of variables” refers to the manipulation of the independent variable and the attempt to maintain the contr
constant value. The method should include explicit reference to how the control of variables is achieved. If the con
practically possible, some effort should be made to monitor the variable(s).
A standard measurement technique may be used as part of a wider investigation but it should not be the focus of t
Students should be assessed on their individual design of the wider investigation. If a standard measurement techn
should be referenced. For example, while planning an investigation to study the effect of light wavelength on the r
in Cabomba, the student may have adapted a method to measure the rate of photosynthesis taken from a textboo
reference would then be expected as a footnote, for example, “Freeland, PW (1985) Problems in Practical Advance
Hodder and Stoughton.” Or the student may adapt a general protocol provided by a teacher in a previous investiga
may appear as: Michigan, J (2007) “Studying the rate of photosynthesis” worksheet.
Students should not be told:
• which apparatus to select
• the experimental method.

Aspect 3: developing a method for collection of data

The definition of “sufficient relevant data” depends on the context. The planned investigation should anticipate the
sufficient data so that the aim or research question can be suitably addressed and an evaluation of the reliability o
made.
If error analysis involving the calculation of standard deviation is to be carried out, then a sample size of at least fi
data range and amount of data in that range are also important. For example, when trying to determine the optim
using a range of pH values between 6 and 8 would be insufficient. Using a range of values between 3 and 10 would
also be insufficient if only three different pH values were tested in that range.
Students should not be told:
• how to collect the data
• how much data to collect.

Data collection and processing

Ideally, students should work on their own when collecting data.
When data collection is carried out in groups, the actual recording and processing of data should be independently
criterion is to be assessed. Recording class or group data is only appropriate if the data-sharing method does not s
presentation format for the students.
Pooling data from a class is permitted where the students have independently organized and presented their data.
may have placed it on a real or virtual bulletin board. For assessment of aspect 1, students must clearly indicate w
own.

Aspect 1: recording raw data

Raw data is the actual data measured. This may include associated qualitative data. It is permissible to convert ha
into word-processed form. The term “quantitative data” refers to numerical measurements of the variables associa
investigation. Associated qualitative data are considered to be those observations that would enhance the interpre
Uncertainties are associated with all raw data and an attempt should always be made to quantify uncertainties. Fo
students say there is an uncertainty in a stopwatch measurement because of reaction time, they must estimate th
uncertainty. Within tables of quantitative data, columns should be clearly annotated with a heading, units and an i
uncertainty of measurement. The uncertainty need not be the same as the manufacturer’s stated precision of the
used. Significant digits in the data and the uncertainty in the data must be consistent. This applies to all measuring
example, digital meters, stopwatches, and so on. The number of significant digits should reflect the precision of th
There should be no variation in the precision of raw data. For example, the same number of decimal places should
derived from processing raw data (for example, means), the level of precision should be consistent with that of the
The recording of the level of precision would be expected from the point where the student takes over the manipul
students would not be expected to state the level of precision in a solution prepared for them.
Students should not be told how to record the raw data. For example, they should not be given a pre-formatted ta
headings, units or uncertainties.

Aspect 2: processing raw data

Data processing involves, for example, combining and manipulating raw data to determine the value of a physical
adding, subtracting, squaring, dividing), and taking the average of several measurements and transforming data in
graphical representation. It might be that the data is already in a form suitable for graphical presentation, for exam
travelled by woodlice against temperature. If the raw data is represented in this way and a best-fit line graph is dra
been processed. Plotting raw data (without a graph line) does not constitute processing data.
The recording and processing of data may be shown in one table provided they are clearly distinguishable.
Students should not be told:
• how to process the data
• what quantities to graph/plot.
Aspect 3: presenting processed data
Students are expected to decide upon a suitable presentation format themselves (for example, spreadsheet, table
diagram, and so on). There should be clear, unambiguous headings for calculations, tables or graphs. Graphs need
scales, labelled axes with units, and accurately plotted data points with a suitable best-fit line or curve (not a scatt
point to data-point connecting lines). Students should present the data so that all the stages to the final result can
Inclusion of metric/SI units is expected for final derived quantities, which should be expressed to the correct numb
figures. The uncertainties associated with the raw data must be taken into account. The treatment of uncertainties
requires the construction of appropriate best-fit lines.
The complete fulfillment of aspect 3 does not require students to draw lines of minimum and maximum fit to the d
error bars or to combine errors through root mean squared calculations. Although error bars on data points (for ex
error) are not expected, they are a perfectly acceptable way of expressing the degree of uncertainty in the data.
In order to fulfill aspect 3 completely, students should include a treatment of uncertainties and errors with their pro
relevant.
The treatment of uncertainties should be in accordance with assessment statements 1.1.2, 1.1.3 and 1.1.4 of this g

Conclusion and evaluation

Aspect 1: concluding
Analysis may include comparisons of different graphs or descriptions of trends shown in graphs. The explanation s
observations, trends or patterns revealed by the data.
When measuring an already known and accepted value of a physical quantity, students should draw a conclusion a
in their result by comparing the experimental value with the textbook or literature value. The literature consulted s
referenced.

Aspect 2: evaluating procedure(s)

The design and method of the investigation must be commented upon as well as the quality of the data. The stude
the weaknesses but must also appreciate how significant the weaknesses are. Comments about the precision and
measurements are relevant here. When evaluating the procedure used, the student should specifically look at the
equipment and management of time.

Aspect 3: improving the investigation

Suggestions for improvements should be based on the weaknesses and limitations identified in aspect 2. Modificat
experimental techniques and the data range can be addressed here. The modifications proposed should be realisti
specified. It is not sufficient to state generally that more precise equipment should be used.

Manipulative skills
(This criterion must be assessed summatively.)

Aspect 1: following instructions

Indications of manipulative ability are the amount of assistance required in assembling equipment, the orderliness
procedure(s) and the ability to follow the instructions accurately. The adherence to safe working practices should b
aspects of practical activities.
A wide range of complex tasks should be included in the scheme of work.

Aspect 2: carrying out techniques

It is expected that students will be exposed to a variety of different investigations during the course that enables t
variety of experimental situations.

Aspect 3: working safely

The student’s approach to safety during investigations in the laboratory or in the field must be assessed. Neverthe
must not put students in situations of unacceptable risk.
The teacher should judge what is acceptable and legal under local regulations and with the facilities available. See
in this guide under “Guidance and authenticity”.
Section A: use of ICT in assessment
Data-logging software may be used in experiments/investigations assessed using the IA criteria provided that the f
applied.
The student’s contribution to the experiment must be evident so that this alone can be assessed by the teacher. T
contribution can be in the selection of settings used by the data-logging and graphing equipment, or can be demon
subsequent stages of the experiment.
(When data logging is used, raw data is defined as any data produced by software and extracted by the student fro
to be subsequently processed by the student.)
The following categories of experiments exemplify the application of this principle.

1. Data logging within a narrowly focused task

Data-logging software may be used to perform a traditional experiment in a new way.
Use of data-logging software is appropriate with respect to assessment if the student decides and inp
relevant software settings. For example, an investigation could be set up to monitor a person’s breathing capa
exercise bike using a spirometer sensor linked to a calculator-based data logger in which the student controls the l
(speed or workload). Data-logging software that automatically determines the various settings and generates the d
graphs would be inappropriate with regard to assessment because the remaining student input required to investig
capacities would be minimal.
If the experiment is suitable for assessment the following guidelines must be followed for the DCP criterion.
Data collection and processing: aspect 1
Students may present raw data collected using data logging as long as they are responsible for the majority of soft
numerical raw data may be presented as a table, or, where a large amount of data has been generated, by graphic
example, the student should set the duration and rate of the sampling, and the generated data in the form of lists
from the calculator or computer could be downloaded by the student into a computer spreadsheet. Students must
correctly, for example, by means of table or graph titles, columns or graph axes labelled with units, indications of u
associated qualitative observations, and so on.
The number of decimal places used in recorded data should not exceed that expressed by the sensitivity of the ins
case of electronic probes used in data logging, students will be expected to record the sensitivity of the instrumen
Data collection and processing: aspects 2 and 3
Use of software for graph drawing is appropriate as long as the student is responsible for most of the decisions, su
•what to graph
•selection of quantities for axes
•appropriate units
•graph title
•appropriate scale
•how to graph, for example, linear graph line and not scatter.
Note: A computer-calculated gradient is acceptable.

In the example of the investigation to monitor breathing capacities, the student could process data by drawing a g
spreadsheet and measuring the breathing frequency from the data. By inspecting the graph or spreadsheet data, t
minimal lung volume values could be identified and used to calculate the mean tidal volume at rest. The mean vol
per minute and recovery rate after exercise could also be calculated.
Statistical analysis carried out using calculators or calculations using spreadsheets are acceptable provided that th
data to be processed and chooses the method of processing. In both cases, the student must show one example in
example, the student must quote the formula used by or entered into a calculator and define the terms used, or th
the formula used in a spreadsheet if it is not a standard part of the program’s menu of functions (for example, mea
deviation).

2. Data logging in an open-ended investigation

Data-logging software can enhance data collection and transform the sort of investigations possible. In this case fu
logging software is appropriate with regard to assessment if it is used to enable a broader, complex investigation t
where students can develop a range of responses involving independent decision-making.
For example, a task could be set to investigate a factor that affects the rate of photosynthesis. If an oxygen sensor
programmed software to monitor the amount of oxygen released by an aquatic plant is used, the student could us
develop a broader, complex investigation, for example, comparing rate of photosynthesis in different species of aq
different light intensities.
Design: aspect 1
The student must state a focused problem/research question, for example: “What is the difference in the rate of ph
different light intensities, as measured by oxygen release, between Elodea canadensis and Myriophyllum spicatum
Relevant variables must also be identified, for example:
• independent variable—species of aquatic plants
• dependent variable—rate of oxygen production
• controlled variables—temperature, mass of plant, leaf surface area, time, light quality
Design: aspect 2
The student must design a method to monitor and control the variables (for example, a water bath for control of te
electronic balance to determine the mass of the plants, and use the same light source to control light quality.
Design: aspect 3
The student must design the method for the appropriate collection of sufficient raw data. The student would select
aquatic plants to use, and measure the amount of dissolved oxygen in the water using the oxygen sensor program
also decide on the range and number of different light intensities and the number of experimental replicates.
Data collection and processing: aspect 1
Appropriate raw data would consist of the rates of photosynthesis derived from the graphs of the experimental run
program using the oxygen sensor. These rates of photosynthesis may be calculated by the student using a function
that analyses the graphs. This must be done without prompting by the teacher. The derived data for rates of photo
annotated on a series of graphs or presented in a table with an appropriate title, column headings and units. Calcu
uncertainties would not be expected in this experiment. In addition, other important data should be recorded, for e
temperature.
Data collection and processing: aspect 2
The graphs showing changes in oxygen concentration would not be assessed, as these would have been generated
the pre-programmed software on the data logger, without input from the student. However, the rates of photosynt
these graphs could be plotted against light intensity for each species using graph-plotting software where student
example, choice of type of graph, x and y axes, range and scale.
Data collection and processing: aspect 3
The student would generate graphs of light intensity versus rates of photosynthesis for each species, which should
correctly labelled axes, a legend for the data of the different species of plants, and trend lines to reveal the degree

Section B: use of ICT in non-assessed practical wor

It is not necessary to use ICT in assessed investigations but, in order to carry out aim 7 in practice, students will be
of the following software applications at least once during the course.
• Data logging in an experiment
• Software for graph plotting
• A spreadsheet for data processing
• A database
• Computer modelling/simulation

Summary of the group 4 project

The group 4 project is a collaborative activity where students from different group 4 subjects work together on a sc
technological topic, allowing for concepts and perceptions from across the disciplines to be shared in line with aim
“encourage an understanding of the relationships between scientific disciplines and the overarching nature of the
The project can be practically or theoretically based. Collaboration between schools in different regions is encourag
The group 4 project allows students to appreciate the environmental, social and ethical implications of science and
also allow them to understand the limitations of scientific study, for example, the shortage of appropriate data and
resources. The emphasis is on interdisciplinary cooperation and the processes involved in scientific investigation, r
products of such investigation.
The choice of scientific or technological topic is open but the project should clearly address aims 7, 8 and 10 of the
guides.
Ideally, the project should involve students collaborating with those from other group 4 subjects at all stages. To th
necessary for the topic chosen to have clearly identifiable separate subject components. However, for logistical rea
may prefer a separate subject “action” phase (see the following “Project stages” section).

Project stages
The 10 hours allocated to the group 4 project, which are part of the teaching time set aside for IA, can be divided i
planning, action and evaluation.

Planning
This stage is crucial to the whole exercise and should last about two hours.
• The planning stage could consist of a single session, or two or three shorter ones.
• This stage must involve all group 4 students meeting to “brainstorm” and discuss the
ideas and information.
• The topic can be chosen by the students themselves or selected by the teachers.
• Where large numbers of students are involved, it may be advisable to have more than
group.
After selecting a topic or issue, the activities to be carried out must be clearly defined before moving
stage to the action and evaluation stages.
A possible strategy is that students define specific tasks for themselves, either individually or as members of group
various aspects of the chosen topic. At this stage, if the project is to be experimentally based, apparatus should be
there is no delay in carrying out the action stage. Contact with other schools, if a joint venture has been agreed, is
consideration at this time.

Action
This stage should last around six hours and may be carried out over one or two weeks in normal scheduled class ti
whole day could be set aside if, for example, the project involves fieldwork.
• Students should investigate the topic in mixed subject groups or single subject groups
• There should be collaboration during the action stage; findings of investigations should
other students within the mixed/single subject group. During this stage, in any practically
important to pay attention to safety, ethical and environmental considerations.
Note: Students studying two group 4 subjects are not required to do two separate action phases.

Evaluation
The emphasis during this stage, for which two hours is probably necessary, is on students sharing their findings, bo
failures, with other students. How this is achieved can be decided by the teachers, the students or jointly.
• One solution is to devote a morning, afternoon or evening to a symposium where all th
individuals or as groups, give brief presentations.
• Alternatively, the presentation could be more informal and take the form of a science f
circulate around displays summarizing the activities of each group.
The symposium or science fair could also be attended by parents, members of the school board and the press. This
pertinent if some issue of local importance has been researched. Some of the findings might influence the way the
its environment or local community.

Addressing aims 7 and 8

Aim 7—“develop and apply the students’ information and communication technology skills in the study of science
Aim 7 may be partly addressed at the planning stage by using electronic communication within and between schoo
(for example, data logging, spreadsheets, databases, and so on) will be used in the action phase and certainly in th
presentation/evaluation stage (for example, use of digital images, presentation software, web sites, digital video, a
Aim 8—“raise awareness of the moral, ethical, social, economic and environmental implications of using science a
The choice of topic should enable one or more elements of aim 8 to be incorporated into the project.

Addressing the international dimension

There are also possibilities in the choice of topic to illustrate the international nature of the scientific endeavour an
cooperation required to tackle global issues involving science and technology. An alternative way to bring an intern
the project is to collaborate with a school in another region.

Types of project
While addressing aims 7, 8 and 10 the project must be based on science or its applications.
The project may have a hands-on practical action phase or one involving purely theoretical aspects. It could be und
range of ways.
• Designing and carrying out a laboratory investigation or fieldwork.
• Carrying out a comparative study (experimental or otherwise) in collaboration with ano
• Collating, manipulating and analysing data from other sources, such as scientific journ
organizations, science and technology industries and government reports.
• Designing and using a model or simulation.
• Contributing to a long-term project organized by the school.

Logistical strategies
The logistical organization of the group 4 project is often a challenge to schools. The following models illustrate pos
the project may be implemented.
Models A, B and C apply within a single school, and model D relates to a project involving collaboration between sc

Model A: mixed subject groups and one topic

Schools may adopt mixed subject groups and choose one common topic. The number of groups will depend on the
The dotted lines in the model show the addition of more groups as student numbers increase.

Model B: mixed subject groups adopting more than one topic

Schools with large numbers of students may choose to do more than one topic.

Model C: single subject groups

For schools opting for single subject groups with one or more topics in the action phase, simply replace the mixed
model A or B with single subject groups.

Model D: collaboration with another school

The collaborative model is open to any school. To this end, the IBO will provide an electronic collaboration board on
schools can post their project ideas and invite collaboration from another school. This could range from merely sha
common topic to a full-scale collaborative venture at all stages.
For schools with few diploma students or schools with certificate students, it is possible to work with non-Diploma P
group 4 students or undertake the project once every two years. However, these schools are encouraged to collab
school. This strategy is also recommended for individual students who may not have participated in the project, fo
illness or because they have transferred to a new school where the project has already taken place.

Timing
The 10 hours that the IBO recommends be allocated to the project may be spread over a number of weeks. The dis
hours needs to be taken into account when selecting the optimum time to carry out the project. However, it is poss
dedicate a period of time exclusively to project work if all/most other school work is suspended.

Year 1
In the first year, students’ experience and skills may be limited and it would be inadvisable to start the project too
However, doing the project in the final part of the first year may have the advantage of reducing pressure on stude
strategy provides time for solving unexpected problems.

Year 1–year 2
The planning stage could start, the topic could be decided upon, and provisional discussion in individual subjects c
end of the first year. Students could then use the vacation time to think about how they are going to tackle the pro
ready to start work early in the second year.

Year 2
Delaying the start of the project until some point in the second year, particularly if left too late, increases pressure
ways: the schedule for finishing the work is much tighter than for the other options; the illness of any student or un
will present extra difficulties. Nevertheless, this choice does mean students know one another and their teachers b
probably become accustomed to working in a team and will be more experienced in the relevant fields than in the

Selecting a topic
Students may choose the topic or propose possible topics, with the teacher then deciding which one is the most vi
resources, staff availability, and so on. Alternatively, the teacher selects the topic or proposes several topics from
a choice.

Assessment
The group 4 project is to be assessed for the personal skills criterion only and this will be the only place whe
assessed. It is up to the school how this assessment takes place.
Personal skills (for group 4 project assessment only)
This criterion addresses objective 4.

Levels/ma Aspect 1 Aspect 2 Aspect 3

rks
Self-motivation and perseverance Working within a team Self-reflection

Complete/ Approaches the project with self- Collaborates and communicates in a Shows a thorough
2 motivation and follows it through to group situation and integrates the own strengths and
completion. views of others. gives thoughtful co
learning experienc

Partial/1 Completes the project but sometimes Exchanges some views but requires Shows limited awa
lacks self-motivation. guidance to collaborate with others. strengths and weak
some consideration
experience.

Not at all/0 Lacks perseverance and motivation. Makes little or no attempt to Shows no awarene
collaborate in a group situation. strengths and weak
consideration to th
experience.

The assessment can be assisted by the use of a student self-evaluation form, but the use of such a form is not a re