Medical applications of processing Flashcards by tcarlso 3

What are the seven steps of tissue processing?

Fixation
Dehydration
Clearing
Embedding
Sectioning
Mounting
Staining

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What is the chemical used in fixation? What does it do?

Formalin or glutaraldehyde

Preserves, fixates, and renders structure resistant to further processing

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What is the chemical used in dehydration? What does it do?

Graded series of EtOH

Removes water from tissue specimen

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What is the chemical used in clearing? What does it do?

Xylene

Prepares the tissue for embedding medium

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What is done in sectioning?

Cut tissue into thin slices

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What is done in mounting?

Place cut section on a slide

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What is the chemical used in staining? Why is staining performed?

Various chemicals are used.

Imparts contrast to tissue so that they can be distinguished

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What is the chemical used in embedding? What does it do?

Paraffin invades tissues, making them hard and section-able

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What is the charge of acid dyes? What structure does this bind to?

negative

mito

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What is the charge of basic dyes? What structure does this bind to?

Positive

Nucleus

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What is eosin?

Acidic, red dye

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What is hematoxylin?

Basic, blue dye

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What is metachromasia?

is a phenomeonon in which a given stain imparts different colors to the tissue (based on density, and wavelength changes)

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What is the term that describes cellular structures that have a net positive charge? Negative?

Acidophilic

Basophilic

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What is the structure affinity (4) and color imparted for hematoxylin?

Nucleus RNA, DNA, ribosomes, a rER

blue

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What is the structure affinity (5) and color imparted for eosin?

Secretory vesicles, sER, lysosomes, mitochondria, type I collagen

pink

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What is the structure affinity and color imparted for feulgen reaction?

DNA magenta

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What is the structure affinity (4) and color imparted for Mallory triple?

Nuceli (red)

muscle (red/orange)

collagen (blue)

Hyaline cartilage (blue)

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What is the structure affinity and color imparted for the PAS reaction?

Carbs

Magenta

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What is the structure affinity and color imparted for the osmic reaction?

Lipids

Black

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What is the structure affinity and color imparted for verhoeff?

Elastic fibers

black

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What is the structure affinity (3) and color imparted for silver methods?

Intermediate filaments of nerve cells, glial cells, and reticular fibers

Black

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What is the structure affinity and color imparted for trypan blue?

macrophages

Blue

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What is the structure affinity and color imparted for prussian blue?

Hemosiderin (ferric Fe)

Blue

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What is the structure affinity and color imparted for Nissl?

Ribosomes Blue

What is the structure affinity (5) and color imparted for Iron hematoxylin?

Nuclear elements, chromosomes, mitochondria, centrioles, and muscle striation Dark blue/black

What are the medical applications of H and E?

coagulative necrosis

What are the medical applications of PAS

identify thickened basement membranes in kidney/glycogen storage diseases and alpha-1 antitrypsin deficiency in liver cells

What are the medical applications of mallory triple?

Highlight fibrosis

What are the medical applications of feulgen reaction?

Demonstrate nuclear changes in CA

What are the medical applications of prussian blue?

excessive Fe accumulation in hemochromatosis

What are the medical applications of congo red?

extracellular deposits of amyloid

What are the medical applications of verhoeff?

elastic fiber in Marfan sydrome

How is necrosis seen when using H and E dyes?

More intense areas of dye uptake (coagulation of cells)

How do you identify increased basement membrane? (what stain, looking for what?)

Use PAS --increased uptake

What is the function of alpha-antitrypsin?

form connective tissue--made in liver Identified by PAS

What stain would you use to identify pulmonary fibrotic lung disease?

Mallory triple (collagen stains)

What stain would you use to look for DNA?

Feulgen reaction

What technique would you use to look for hemochromotosis?

Prussian blue

What stain would you use to identify amyloid?

Congo red

What chemical would you use to identify elastic fibers?

Verhoeff

When are frozen tissue sections used?

Useful for urgent diagnosis (but detail lost)

What is immunocytochemistry? When is it utilized?

Using labeled antibodies Estrogen receptors, Categorize CA cells

What is the difference between the direct versus the indirect method of immunocytochemistry?

Direct = antigen and marker are one Indirect = antigen separate from marker--marker added to another antibody which attaches to the first antibody

What is a more accurate method of detection, direct or indirect immunocytochemisty?

Indirect (more markers)

What method would you use to categorize a tumor cell with cytokeratin/epithelial cells?

Immunocytochemistry

What is kaposi sarcoma? What antigen do they express?

CA of the lymphatic epithelium D2-40

How would you identify a kaposi sarcoma under a light microscope?

Apply tagged antibodies to D2-40

What type of method would you use to identify HER2/Neu receptors or estrogen receptors?

Immunocytochemistry

What is In situ hybridization?

Instead of antibodies, this technique uses complementary nucleic acid probes to specifically identify a nuclei acid sequence of interest

Why would you use in situ hybridization?

Identify a specific virus | Detect gene amplification

What type of molecule is heparin? Where is it found?

GAG mast cells

Is heparin basophilic or acidophilic? Why?

Basophilic, has negatively charge sulfates

Is toludine blue an acidic or basic dye? What color is it?

basic blue

In alpha-1-antitrypsin disease, where does misfolded trypsin accumulate? What are the symptoms?

Proteins accumulate in the sER. COPD (even w/o smoking)

What color does Mallory triple stain Nuclei?

red

What color does Mallory triple stain muscle cells?

red

What color does Mallory triple stain collagen?

Blue

What color does Mallory triple stain hyaline cartilage?

Blue

What color will congo red appear in normal light? Polarized light?

Red in normal light | Red-green in polarized light

What type of stain would you use to highlight elastic fibers like in Marfan syndrome?

Verhoeff

What is immunocytochemistry used in diagnosing CA?

To see where the tumor originated from, and how aggressive it is. To see if ER+

anticytokeratin comes from what tissue?

Epithelial

Kaposi sarcoma is CA derived from what tissue type?

Lymphatic cells

What is the specific antigen marker that lymphatic vessels express?

D2-40

You are trying to determine if a patient has the BRCA1/2 gene. What is one way to determine if they have this mutation?

FISH

Overexpression of ALK (a Y kinase) is indicative of what?