specimen assays
DESCRIPTION
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SPECIMEN ASSAYS
Yenny Yustisia
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Surface characterization
Contact angle analysis
Light microscopy
Electron microscopy
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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
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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
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Bright Field (Khler illumination) Microscopy
Dark Field Microscopy
Phase Contrast
Polarised Light Microscopy
Differential Interference Contrast
Fluorescence Microscopy
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Confocal Microscopy
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Electron microscopy
a microscope that uses accelerated electronsas a source of illumination
Scanning Electron Microscopy (SEM)
Transmission Electron Microscopy (TEM)
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CR
T
Detector
Amplifier
Electron gun
Condenser lens
Objective lens
Magnification control unit
Scanning circuit
Beam deflector
Specimen
Condenser lens
Specimen
Objective lens
Lampu
Intermediate lens
Projector lens
Projector screen
Penyinaran dan pembentukan bayangan pada MO, TEM, dan SEM
TEM SEMMO
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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)
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(a) Wetting system showing forces acting on the liquid drop.
(b) Nonwetting system with > 90 .
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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.
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Four possibilities for contact angle measurement: (A) sessile drop, (B) captive air bubble method, (C) capillary rise method, (D) Wilhelmy plate method.
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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
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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
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Assays for protein type and amount
High-Performance Liquid Chromatography (HPLC)
Colorimetric assays
Enzyme-linked Immunosorbent Assay (ELISA)
Western Blotting
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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 solventcontaining 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
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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
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Fluorescent assays
Similar in concept to colorimetric assays
Key difference: reaction causes the attachment of a fluorescing molecule (fluorophore) to the protein of interest
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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
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Western Blotting
Assay to identify a certain protein after separation of the many proteins contained in a sample through the use of gel electrophoresis
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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
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DNA and RNA assays
To determine gene mutation, damaged gene
Polymerase Chain Reaction (PCR)
Southern and Northern Blotting
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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) DNAReverse-Transcription PCR RNA