Yenny Yustisia

SPECIMEN ASSAYS

Surface characterization
Contact angle analysis
Light microscopy
Electron microscopy

Light microscopy
an instrument that uses visible light and
magnifying lenses to examine small objects

not visible to the naked eye, or in finer detail

than the naked eye allows

techniques
Transmitted Light Microscopy
is the general term used for any

type of microscopy where the light

is transmitted from a source on
the opposite side of the specimen
to the objective lens

Inverted
observing living cells or organisms

at the bottom of a large container

(e.g., a tissue culture flask) under
more natural conditions than on a
glass slide

 Bright Field (Khler illumination)

Microscopy
Dark Field Microscopy
Phase Contrast
Polarised Light Microscopy
Differential Interference Contrast

Fluorescence Microscopy

 Confocal Microscopy

Electron microscopy
a microscope that uses accelerated electrons
as a source of illumination
Scanning Electron Microscopy (SEM)
Transmission Electron Microscopy (TEM)

Penyinaran dan pembentukan bayangan pada MO, TEM, dan SEM

Lampu
Electron gun
Scanning circuit

Condenser
lens

Condenser
lens
Specimen

Magnification
control unit

Objective lens
Beam
deflector

Objective
lens

Detector

Intermediate
lens
Projector lens

Amplifier
Specimen

CRT

Projector
screen

MO

TEM

SEM

Contact angle analysis

The force balance between the liquidvapor

surface tension (lv) of a liquid drop and the

interfacial tension between a solid and the drop
(sl), manifested through the contact angle () of
the drop with the surface, can be used to
quantitatively characterize the energy of the
surface (sv)

(a) Wetting system showing forces acting on the liquid drop.

(b) Nonwetting system with > 90 .

The basic relationship describing this force balance is:

sv = sl + lvcos

The energy of the surface, which is directly related to its

wettability, is a useful parameter that has often correlated
strongly with biological interaction.

Four possibilities for contact angle measurement: (A) sessile

drop, (B) captive air bubble method, (C) capillary rise method, (D)
Wilhelmy plate method.

Spectrophotometry
to measure the amount of light that a sample absorbs.

The instrument operates by passing a beam of light

through a sample and measuring the intensity of light
reaching a detector

Infrared Spectroscopy
a simple and reliable

technique widely used in

both organic and
inorganic chemistry
A Molecul was indentifed
by its molecular vibration,
based on the absorbtion
and intensity of spesific
infrared wavelengths
Provides information on
functional groups

Assays for protein type and

amount
High-Performance Liquid Chromatography
(HPLC)
Colorimetric assays

Enzyme-linked Immunosorbent Assay

(ELISA)
Western Blotting

High-Performance Liquid
Chromatography (HPLC)
is a technique in analytic chemistry used to separate the

components in a mixture, to identify each component, and

to quantify each component
It relies on pumps to pass a pressurized liquid solvent
containing the sample mixture through a column filled with
a solid adsorbent material
Each component in the sample interacts slightly differently
with the adsorbent material, causing different flow rates for
the different components and leading to the separation of
the components as they flow out the column
Ex: detecting vitamin D levels in blood serum, separating
the components of a complex biological sample, or of
similar synthetic chemicals from each other

Colorimetric assays
Assays for the presence of a certain protein,
based on changes in an observable quantity

such as color
applicable to both organic compounds and
inorganic compounds and may be used with
or without an enzymatic stage

Fluorescent assays
Similar in concept to colorimetric assays
Key difference: reaction causes the

attachment of a fluorescing molecule

(fluorophore) to the protein of interest

Enzyme-linked Immunosorbent
Assay (ELISA)
a test that uses antibodies and color change
to identify a substance
For a very specific identification of a protein

Western Blotting
Assay to identify a certain protein after
separation of the many proteins contained in

a sample through the use of gel

electrophoresis

Immunostaining
To identify the location of proteins in tissues
and visualized via light microscopy
A primary antibody is added to the section bind

protein of interest
Second Ab linked to and anzyme or chromophore
binds primary Ab
Imaged under visible light

DNA and RNA assays

To determine gene mutation, damaged gene
Polymerase Chain Reaction (PCR)

Southern and Northern Blotting

Polymerase Chain Reaction (PCR) &

Reverse-Transcription PCR
To expand the amount of DNA or RNA for
that gene to detectable levels
Polymerase Chain Reaction (PCR) DNA
Reverse-Transcription PCR RNA