Fixation - ANS >> Treatment of tissue with chemical agents that retard alterations of the tissue subsequent to death but also maintain normal structure
Formalin and Bouin's fluid are the most common fixative agents
Dehydration - ANS >> A graded series of alcohol baths 50% -100...
GMS 6630 Histology USF Questions Updated To
Pass
Fixation - ANS >> Treatment of tissue with chemical agents that retard alterations of the tissue
subsequent to death but also maintain normal structure
Formalin and Bouin's fluid are the most common fixative agents
Dehydration - ANS >> A graded series of alcohol baths 50% -100% are used to remove the water.
The tissue is then treated with xylene a chemical that is miscible with melted paraffin
Sectioning - ANS >> Blocks of tissue are then sliced into thin segments about 5-10 um. Also can be
performed on frozen samples (liquid nitrogen)
Mounting and staining - ANS >> Paraffin sections are mounted on glass slides and then stained by
water soluble stains that permit differentiation of the various components.
The paraffin is removed and the section is stained. The section is then dehydrated again and placed
under a coverslip.
Light microscopes - ANS >> Use an arrangement of lenses to magnify an image
Compound microscope (uses more than one lense.
The light source is a light bulb whose light is gathered into a focus via a condenser lense.
Image from the objective lens is gatherers and further magnified by the eyepiece
This combined with the ocular lens leads to magnification of 40 100 400 and 1000
Quality of the image is referred to as resolution- ability of the lens to show that two objects are
separated by a distance
`
Digital imaging tech - ANS >> Allows immediate visualization of the acquired image
Digital modification of the image
Capability of enhancing the image by the use of commercially available software
, Interpretation of microscopic section - ANS >> Most difficult part is determining the #D structure via 2d
images.
Advanced Visualization - ANS >> Histochemistry
Immunochemistry
Confocal microscopy
Electron microscopy
Histochemistry - ANS >> Method of staining tissue that provides information about the presence and
location of intracellular and extracellular macromolecules
Specific chemical chemical constituents of tissues and cells can be localized by the methods of
histochemistry and cytochemistry. These reactions capitalize on the enzyme activity, chemical reactivity,
and other physicochemical phenomena associated with the constituent of interes
immunocytochemistry (ICC) - ANS >> uses fluoresceinated antibodies and anti-antibodies to provide
more precise intracellular and extracellular localization of macromolecules than is possible with
histochemistry.
Antibody labelling - ANS >> Direct and indirect methods
Direct Antibody labelling - ANS >> Antibody bound to macromolecule is labelled with a fluorescent dye.
The antibody is then permitted to react with the macromolecule and the resultant complex may be
viewed with a fluorescent microscope
indirect method - ANS >> A fluorescent labelled antibody is prepared against the primary antibody
bound to the macromolecule
Once the primary antibody has been bound the area is washed to remove excess, the labelled antibody
is then added and forms a complex visible by fluorescent microscopy
The indirect method is more visible because multiple secondary antibodies will bind to the primary
making them more sensitive.
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