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18 Cards in this Set

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What can be used to fix a kidney biopsy for EM?

Initial fixation in NBF, then glutaraldehyde.


Or Carson-Millonig buffered formalin.


Or initial fix in glutaraldehyde.

For EM, if primary fixative is glutaraldehyde, the second fixative is usually what?

Osmium teroxide, which fixes lipids sufficiently.

Which method of enzyme activity histo chemistry can show different types of WBC?

Chloroacetate esterase.

Which enzyme activity histochemistry can show different type of fibers in muscle?

ATPase

Enzymes commonly used in IHC to unmask antigenic determinants?

Pronase, trypsin, proteinase K, pepsin.

What are the tissues most often tested for diagnostic using fluorescent-labeled antibodies?

Skin and kidney

Sections used for EM are cut at what thickness?

0.05 microns. Ultra structural examination are cut at 100 to 800 angstrom (10,000 angstrom equals 1000 nanometers)

In order to color detect reaction product, peroxidase does what reaction to hydrogen peroxide?

HRP catalyzes the release of oxygen from hydrogen peroxide (that is, reduction), which is coupled with the oxidation of a chromogenic substrate.

What is the preferred fixative for intermediate filaments?

Absolute ethanol or an alcohol based fixative.

Describe the chemicals reaction that forms the visible product in the cobalt-calcium method for ATPase.

The final product, calcium phosphate, is visualized by first adding cobalt chloride, which converts it into cobalt phosphate. Then adding ammonium or sodium sulfide turns cobalt phosphate into cobalt sulfide, which is black. The goal is to turn calcium phosphate into cobalt sulfite, which is black.

When would the chromogenic substrate amino-ethylcarbazole (AEC) be preferential to DAB?

When staining pigmented lesions. Because AEC results in pink to red color, it won't be confused with melanin; ie brown like DAB.

Which chromogen used in peroxidase histochemistry generates a product that is soluble in alcohol or xylene but not water.

9-amino-ethylcarbazole. DAB is insoluble in any solvents.

What is the chemical reaction that forms the secondary reaction product (often insoluble) in nearly all histochemical methods for enzyme activity?

Trapping, which includes reduction reactions.

Which type of cells are responsible for immunoglobulin production?

Plasma cells, which are fully differentiated B-lymphocytes.

When ATPase activity is measured in muscle, what component of the muscle is actually being measured?

Myosin ATPase. Note sodium pump ATPase activity of cell membrane is not measured because histochemical medium doesn't provide such suitable condition.

Is sodium sulfite considered to be a toxic or carcinogen hazard?

Yes

What is a myeloma cell and how is it used is hybridoma technology (in antibody production)?

Myeloma cells are immortal, malignant plasma cells. The spleen cells of an animal that is immunized with a specific antigen is fuse with myeloma cells. The single hybrid cell which produce IgGs (spleen) and immortal (myeloma) can be cultured to produce large amounts of specific antibody.

An antibody against which ubiquitous intermediate filament is typically included to assess over fixation.

Vimentin, which is expressed by mesenchymal cells present in all tissues.