How to Use Microscope and Cell Observation

Okty Chairunnisa Wangsa Praja

3315115798


FMIPA UNJ
2011





Title
How to use microscope and observe the cells

B Purpose
1. Learn how to prepare the materials that will be observed under the microscope.
2. Learn how to use microscope correctly.
3. Observe living cell and coati cell.
4. Observe the cell shapes.
5. Observe the diIIerences between plant cell and animal cell.

C Theoritical Background

Microscope
The microscope that we use is a Light Microscope. This microscope works
when light can through the specimen and penetrate the glass lens. This lens reIracting
the light so the specimen shadow was magniIying when that shadows projected to our
eyes.


The importance things that you should considered are:
1. Hold the microscope tightly in the arm with one hand and the other hand to
prop the microscope.
2. Use the microscope with the arm in Iront oI us.
3. Put the microscope on sturdy table. Don`t put the microscope on book or piles
oI paper.
4. Object table must be horizontal so preparat not Ialls.
5. Clean the lens only with lens paper.
6. II themicroscope using the light, don`t move and leave the microscope on.
And keep the microscope when the temperature goes down.
7. Both oI our eyes should be opened when observing with microscope.


How to Use The Microscope :
First, think about what you want to do with the microscope. What is the maximum
magniIication you will need? Are you looking at a stained specimen? How much
contrast/resolution do you require? Next, start setting up Ior viewing.
1 ount the specimen on the stage
The cover slip must be up iI there is one. High magniIication objective lenses can't
Iocus through a thick glass slide; they must be brought close to the specimen, which is
why coverslips are so thin. The stage may be equipped with simple clips (less
expensive microscopes), or with some type oI slide holder. The slide may require
manual positioning, or there may be a mechanical stage (preIerred) that allows precise
positioning without touching the slide.
2 Optimize the lighting
A light source should have a wide dynamic range, to provide high intensity
illumination at high magniIications, and lower intensities so that the user can view
comIortably at low magniIications. Better microscopes have a built-in illuminator,
and the best microscopes have controls over light intensity and shape oI the light
beam. II your microscope requires an external light source, make sure that the light is
aimed toward the middle oI the condenser. Adjust illumination so that the Iield is
bright without hurting the eyes.
3 djust the condenser
To adjust and align the microscope, start by reading the manual. II no manual is
available, try using these guidelines. II the condenser is Iocusable, position it with the
lens as close to the opening in the stage as you can get it. II the condenser has
selectable options, set it to bright Iield. Start with the aperture diaphragm stopped
down (high contrast). You should see the light that comes up through the specimen
change brightness as you move the aperture diaphragm lever.
4 Think about what you are looking for
It is a lot harder to Iind something when you have no expectations as to its
apprearance. How big is it? Will it be moving? Is it pigmented or stained, and iI so
what is its color? Where do you expect to Iind it on a slide? For example, students
typically have a lot oI trouble Iinding stained bacteria because with the unaided eye
and at low magniIications the stuII looks like dirt. It helps to know that as smears dry
down they usually leave rings so that the edge oI a smear usually has the densest
concentration oI cells.
5 Focus, locate, and center the specimen
Start with the lowest magniIication objective lens, to home in on the specimen and/or
the part oI the specimen you wish to examine. It is rather easy to Iind and Iocus on
sections oI tissues, especially iI they are Iixed and stained, as with most prepared
slides. However it can be very diIIicult to locate living, minute specimens such as
bacteria or unpigmented protists. A suspension oI yeast cells makes a good practice
specimen Ior Iinding diIIicult objects.
O Use dark Iield mode (iI available) to Iind unstained specimens. II not, start with high
contrast (aperture diaphragm closed down).
O Start with the specimen out oI Iocus so that the stage and objective must be brought
closer together. The Iirst surIace to come into Iocus as you bring stage and objective
together is the top oI the cover slip. With smears, a cover slip is Irequently not used,
so the Iirst thing you see is the smear itselI.
O II you are having trouble, Iocus on the edge oI the cover slip or an air bubble, or
something that you can readily recognize. The top edge oI the cover slip comes into
Iocus Iirst, then the bottom, which should be in the same plane as your specimen.
O Once you have Iound the specimen, adjust contrast and intensity oI illumination, and
move the slide around until you have a good area Ior viewing.
6 djust eyepiece separation, focus
With a single ocular, there is nothing to do with the eyepiece except to keep it clean.
With a binocular microscope (preIerred) you need to adjust the eyepiece separation
just like you do a pair oI binoculars. Binocular vision is much more sensitive to light
and detail than monocular vision, so iI you have a binocular microscope, take
advantage oI it.
One or both oI the eyepieces may be a telescoping eyepiece, that is, you can Iocus it.
Since very Iew people have eyes that are perIectly matched, most oI us need to Iocus
one eyepiece to match the other image. Look with the appropriate eye into the Iixed
eyepiece and Iocus with the microscope Iocus knob. Next, look into the adjustable
eyepiece (with the other eye oI course), and adjust the eyepiece, not the microscope.
7 Select an objective lens for viewing
The lowest power lens is usually 3.5 or 4x, and is used primarily Ior initially Iinding
specimens. We sometimes call it the scanning lens Ior that reason. The most
Irequently used objective lens is the 10x lens, which gives a Iinal magniIication oI
100x with a 10x ocular lens. For very small protists and Ior details in prepared slides
such as cell organelles or mitotic Iigures, you will need a higher magniIication.
Typical high magniIication lenses are 40x and 97x or 100x. The latter two
magniIications are used exclusively with oil in order to improve resolution.
Move up in magniIication by steps. Each time you go to a higher power objective, re-
Iocus and re-center the specimen. Higher magniIication lenses must be physically
closer to the specimen itselI, which poses the risk oI jamming the objective into the
specimen. Be very cautious when Iocusing. By the way, good quality sets oI lenses
are parIocal, that is, when you switch magniIications the specimen remains in Iocus or
close to Iocused.
Bigger is not always better. All specimens have three dimensions, and unless a
specimen is extremely thin you will be unable to Iocus with a high magniIication
objective. The higher the magniIication, the harder it is to "chase" a moving specimen.
djust illumination for the selected objective lens
The apparent Iield oI an eyepiece is constant regardless oI magniIication used. So it
Iollows that when you raise magniIication the area oI illuminated specimen you see is
smaller. Since you are looking at a smaller area, less light reaches the eye, and the
image darkens. With a low power objective you may have to cut down on illumination
intensity. With a high power you need all the light you can get, especially with less
expensive microscopes.
9 When to use bright field microscopy
Bright Iield microscopy is best suited to viewing stained or naturally pigmented
specimens such as stained prepared slides oI tissue sections or living photosynthetic
organisms. It is useless Ior living specimens oI bacteria, and inIerior Ior non-
photosynthetic protists or metazoans, or unstained cell suspensions or tissue sections.
Here is a not-so-complete list oI specimens that might be observed using bright-Iield
microscopy, and appropriate magniIications (preIerred Iinal magniIications are
emphasized).
O Prepared slides, stained - bacteria (1000x), thick tissue sections (100x, 400x), thin
sections with condensed chromosomes or specially stained organelles (1000x), large
protists or metazoans (100x).
O Smears, stained - blood (400x, 1000x), negative stained bacteria (400x, 1000x).
O Living preparations (wet mounts, unstained) - pond water (40x, 100x, 400x), living
protists or metazoans (40x, 100x, 400x occasionally), algae and other microscopic
plant material (40x, 100x, 400x). Smaller specimens will be diIIicult to observe
without distortion, especially iI they have no pigmentation.



Cells
Cells are the 'Building Blocks oI liIe. Most oI cells are so small so you need a microscope
to see them. Cells were divided into two, there are prokaryotic and eukaryotic. Prokaryotic
are bacteria and archaea. Eukaryotic are plant, Iungi and animal.

Plant Cell
Plants are unique among the eukaryotes, organisms whose cells have membrane-enclosed
nuclei and organelles, because they can manuIacture their own Iood. Chlorophyll, which
gives plants their green color, enables them to use sunlight to convert water and carbon
dioxide into sugars and carbohydrates, chemicals the cell uses Ior Iuel.
Like the Iungi, another kingdom oI eukaryotes, plant cells have retained the protective cell
wall structure oI their prokaryotic ancestors. The basic plant cell shares a similar construction
motiI with the typical eukaryote cell, but does not have centrioles, lysosomes, intermediate
Iilaments, cilia, or Ilagella, as does the animal cell. Plant cells do, however, have a number oI
other specialized structures, including a rigid cell wall, central vacuole, plasmodesmata, and
chloroplasts. Although plants (and their typical cells) are non-motile, some species produce
gametes that do exhibit Ilagella and are, thereIore, able to move about.










Animal Cells
Inside the animal cells there are so much component called organel. The organel was veiled
by membrane. Animal cells are like tiny, jelly-Iilled sacs. Each sac has a skin like cover
called a cell membrane, which encloses a jelly-like substance called cytoplasm within the
cell. Organelles in the cells do the diIIerent jobs. The nucleus is the important organelle. The
nucleus is the cells 'brain. It is the control center Ior all the diIIerent jobs that take place
inside the cell. The nucleus also contains instructions that tell new cells how to grow and
Iunction.
















Golgi pparatus: A series (stack) oI Ilattened, membrane-bound sacs (saccules) involved
in the storage, modiIication and secretion oI proteins (glycoproteins) and lipids destined to
leave the cell (extracellular) and Ior use within the cell (intracellular). The Golgi apparatus is
abundant in secretory cells, such as cells oI the pancreas.
Golgi Vesicle: A membrane-bound body that Iorms by "budding" Irom the Golgi
apparatus. It contains proteins (glycoproteins), such as digestive enzymes, and migrates to the
cell (plasma) membrane. Golgi vesicles Iuse with the cell membrane and discharge their
contents into the exterior oI the cell through a process called exocytosis. Some Golgi vesicles
become lysosomes which are involved in intracellular digestion.
Pinocytotic Vesicle: A membrane-bound vacuole Iormed by a speciIic type oI
endocytosis called pinocytosis. The plasma membrane invaginates (pinches inwardly) to Iorm
a vesicle that detaches and moves into the cytoplasm. Macromolecular droplets and particles
up to 2 micrometers in diameter enter the cell within these pinocytotic vesicles. Larger
particles (including bacteria) enter special white blood cells (phagocytes) through a Iorm oI
endocytosis called phagocytosis. The moeba is a unicellular protist that ingests Iood
(including algal cells) by phagocytosis.
Lysosome: A membrane-bound organelle containing hydrolytic (digestive) enzymes.
Lysosomes originate as membrane-bound vesicles (called Golgi vesicles) that bud Irom the
Golgi apparatus. They are primarily involved with intracellular digestion. Lysosomes Iuse
with vesicles (small vacuoles) Iormed by endocytosis. The contents oI these vesicles are
digested by lysosomal enzymes. Autodigestion by lysosomes also occurs during embryonic
development. The Iingers oI a human embryo are webbed initially, but are separated Irom
each other by lysosomal enzymes. Cells in the tail oI a tadpole are digested by lysosomal
enzymes during the gradual transition into a Irog.
Peroxisome: A membrane-bound organelle that contains speciIic enzymes imported Irom
the cytoplasm (cytosol). For example, certain peroxisomes contain the enzyme catalase which
rapidly breaks down toxic hydrogen peroxide into water and oxygen. This reaction can be
easily demonstrated by pouring some hydrogen peroxide on raw meat or an open wound.
Glycolysis: An anaerobic oxidation pathway outside oI the mitochondria in which glucose
is oxidized to pyruvate with a net gain oI 2 ATP molecules. Pyruvate is converted into a 2-
carbon acetyl group which enters the Krebs cycle within the mitochondria.
itochondrion: Membrane-bound organelle and the site oI aerobic respiration and ATP
production. Energy Irom the step-by-step oxidation oI glucose (called the Krebs or citric acid
cycle) is used to produce molecules oI adenosine triphosphate (ATP). The Krebs cycle starts
when a 2-carbon acetyl group combines with a 4-carbon group to Iorm a 6-carbon citrate.
Including glycolysis (which occurs outside the mitochondria), a total oI 38 ATP molecules
are generated Irom one molecule oI glucose.
Centrioles : Nonmembrane-bound organelles that occur in pairs just outside the nucleus oI
animal cells. Each centriole is composed oI a cylinder or ring oI 9 sets oI microtubule triplets
with none in the middle (9 0 pattern). During cell division a pair oI centrioles moves to
each end oI the cell, Iorming the poles oI the mitotic spindle. Centrioles also give rise to basal
bodies that control the origin oI cilia and Ilagella in motile cells oI protists. In cross section,
Ilagella and cilia have 9 sets oI microtubule doublets surrounding a pair oI single
microtubules in the center (9 2 pattern). This characteristic pattern also occurs in motile
cells oI higher organisms, such as human sperm.

The DiIIerences between Plant Cell and Animal Cell
Animal cells do not have a cell wall. Instead oI a cell wall, the plasma membrane (usually
called cell membrane when discussing animal cells) is the outer boundary oI animal cells.
Animal tissues thereIore require either external or internal support Irom some kind oI
skeleton. Frameworks oI rigid cellulose Iibrils thicken and strengthen the cell walls oI higher
plants. Plasmodesmata that connect the protoplasts oI higher plant cells do not have a
counterpart in the animal cell model. During telophase oI mitosis, a cell plate is Iormed as
the plant cell begins its division. In animal cells, the cell pinches in the center to Iorm two
cells; no cell plate is laid down. Centrioles are generally not Iound in higher plant cells,
while they are Iound in animal cells. Animal cells do not have plastids, which are common in
plant cells (chloroplasts). Both cell types have vacuoles, however, in animal cells vacuoles
are very tiny or absent, while in plant cells vacuoles are generally quite large.







D aterials and ethod

Apparatus :
1. Light microscope
2. Object glass
3. Cover glass
4. Cutter
5. Tissue
6. Toothpicks

Materials :
1. piece oI paper bearing the A letters
2. Dried shallot skin (Album cepa)
3. Adam and Eva Leaves (#4e4 disc4l47)
4. Hydrilla (d7illa ve79icila9a)
5. Air plant (alanc4e pinna9a)
6. Cheek mucosal ephitelium
Method :
Activity 1 : Practice Using The Microscope





Activity 2 : Observe Coati Cell in Plant
Activity 3 : Observe the Iresh preparat Irom cell plant
a. Adam and Eva leaves (Rhoeo discolor)


b. H
y
LeLLer A
pleces
puL ln
ob[ecL glass
+ waLer
cover wlLh
Lhe cover
glass
observe
wlLh
mlcroscope
drled shalloL
skln
puL ln ob[ecL
glass +
waLer
dropplng
Lhe PCL
cover wlLh
Lhe cover
glass
observe
wlLh
mlcroscope
underleaves
eplderm
puL ln Lhe
ob[ecL glass +
waLer
cover wlLh
cover glass
observe wlLh
Lhe
mlcroscope
b. Hydrilla (Hydrilla verticillata)


c. Air Plant (Kalanchoe pinnata)

d. Observe the Iresh preparat Irom animal cell



Lear Lhe
hydrllla
puL ln Lhe
ob[ecL glass
+ waLer
cover wlLh
Lhe cover
glass
observe lL
wlLh Lhe
mlcroscope
sllce Lhe
leave
eplderm
puL ln Lhe
ob[ecL glass
+ waLer
cover wlLh
Lhe cover
glass
observe
wlLh
mlcroscope
cheek
ephlLel
puL ln Lhe
ob[ecL glass
+ waLer
cover wlLh
Lhe cover
glass
observe
wlLh Lhe
mlcroscope
E #esult and Discussion

Result

A. Adam and Eve LeaI (#4ea disc4l47)

InIormation :
A. MagniIication 4 x 10 40
B. InIormation oI the Iigure :
1. Wall Cell
2. Cytoplasm
3. Stomata


B. Hydrilla (d7illa ve79icila9a)

InIormation :
A. MagniIication 4 x 10 40
B. InIormation oI the Iigure
1. A liquid comes Irom
cytoplasm (cell)
2. Wall Cell
3. Chloroplast


C. Air Plant (alanc4e pina9a)

InIormation :
A. MagniIication 4 x 10 40
B. InIormation oI the Iigure
1. Wall Cell
2. Stomata
3. Space between cell

C. Observing Animal Cell
InIormation :
A. MagniIication 4 x 10 40
B. InIormation oI the Iigure
1. Membrane Cell
2. Nucleus
3. Cytoplasm




Discussion
Activity 1 : Practice Using the Microscope
AIter haved an observation in A letter titled paper on the glass object and then
observed with weak magniIying, the A shadow was reverse. Because the objective
lens and eyepiece are both convex lens. Broad objective lens produces a temporary reIlection
oI a Iictitious character, upside-down and zoomed. Then to determine the character oI
terminal reIlection is okuler lens. In light microscope, the terminal reIlection have the same
characteristic as a temporary reIlection, apparent, reverse and more enlarged.
Activity 2 : Observe the Coati Cell in plant
In Iigure, there is a cell wall and cytoplasm. Inside the cell oI shallot there is a
useIul cell wall to maintain the Iorm oI cells and protect cells Irom mechanical
damage and there is a Ieature that the cell walls oI shallot skin cell is a plant cell.
In the shallot cell experiment, we use the HCl because acidic concentrated HCl can
damage the nucleus and the nucleus eventually mixes with the cytoplasm. So we can`t see the
cytoplasm. The destruction oI nucleus make us can`t see the other organelle because nucleus
is the center regulator oI cell.
Activity 3 : Observe the Iresh preparat Irom cell plant
a. Adam and Eva leaves (#4e4 disc4l47)
In Iigure there are a cell wall, cytoplasm, and stomata. In the adam and eva cell there
is a useIul cell wall to maintain the Iorm oI cells and protect cells
Irom mechanical damage and the Iunction oI cell wall was a characteristic that Adam
and Eva is a plant cell.


b. Hydrilla (d7illa ve79icilla9a)
In Iigure there are a cell wall, cytoplasm, and stomata. Same with Adam and Eva and
shallot cell, inside the hydrilla there`s a cell wall to maintain the Iorm oI cells and
protect cells Irom mechanical damage and the Iunction oI cell wall was a
characteristic that Hydrilla is a plant cell. Cytoplasm is a protoplasm part in a jelly
Iorm. Most oI the cytoplasm are water and a little bit oI koloid. Chloroplast was
shaped like a lens, usually the size was 4-6 m. Chlorophyll contained in chloroplast.

c. Air Plant (alanc4e pinna9a)
In Iigure there are a cell wall, cytoplasm, and stomata. Same with Adam and Eva,
shallot cell and Hydrilla, inside the Air Plant there`s a cell wall and it shows that Air
plant is a plant cell. In Air plant there`s a middle lamela. Only Air Plant have a middle
lamela that`s because Air Plant lives on top oI water so it needs a lower density than
water to Iloating.

d. Observe the Iresh preparat Irom animal cell
In the experiment oI animal cell we use human cheek mucosa ephitelium. In Iigure,
there are cell membrane, cytoplasm and nucleus. There`s no cell wall, it`s because the
human epitel not a plant cell. The lack oI cell wall in animal cells involve the cells in
animal cells are irregular shape. Cell membrane was a transportation instrument Ior
cell. Cell membrane is also a means oI transportation Ior the cell entry and exit
oI substances that are needed and not needed by the cell.The components
oI cell membranes include phospholipids, proteins, oligosaccharides,
glycolipids and cholesterol.

F #eferences
99p.//www.7uf.7ice.edu/~bi4slabs/me94ds/mic74sc4p/mic74sc4p.9ml
99p.//www.mic74.magne9.fsu.edu/
Silve7s9ein, Alvin., Silve7s9ein, Ji7ginia., Silve7s9ein, Ji7ginia B., Silve7s9ein, Lau7a Nun.
2009. Cells.Twen9i7s9 Cen9u7 B44s.
G7een, Jen. 2010. Inside Animals. Marshall Cavendish.
99p.//wanesw47d.pal4ma7.edu/lmexe71a.9ml




G Suplement
1. Compare the location oI the image with the location oI the object being observed. Whether
the shadow lies the same or inverted? Whether the reIlection is a mirror reIlection?
- Inve79ed. N4, i9s a lens 7eflec9i4n.
2. While looking at the okuler lens, slide the preparations Irom leIt to right. Which way
the shadows move? Which way the shadow shiIts iI preparations are shiIted Iorwards?
- the shadows move to the left. If the preparations was shifted forwards so the
shadow will be rearward.
3. Rotate the weak objective to the strong objective. Whether that rotate changing
the view to be broad or narrow? Whether the replacement is changing the reflection
position? And whether the reflection looks darker or lighter?
- it looks broad. No, the reflection still looks inverted and darker.