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Histology > Tissue Prep/Staining > Flashcards

Tissue Prep/Staining Flashcards

1
Q

Fixing

A
  • prevents deterioration, and hardens tissue.
  • formalin is most common: reacts with amino acid to stabilize tissue structure, not good for detail
  • all contain glacial acetic acid: counters shrinkage
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2
Q

Basic Fixing

A
  • used for mitochondrial staining.

- chromatin is dissolved

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3
Q

Why must sample be dehydrated?

A

-will be embedded with paraffin (hydrophobic material)

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4
Q

How is a sample dehydrated?

A
  • series of more concentrated ethanol baths

- will destroy neutral fats

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5
Q

What happens when a sample is cleared?

A

-alcohol is replaced with xylene or cedar oil

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6
Q

What does clearing do?

A

Removes the paraffin embedded medium.

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7
Q

List three clearing agents.

A
  • xylene
  • cedar oil
  • carbon tetrachloride
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8
Q

What is embedding process?

A
  • specimen moved through three melted paraffin baths.
  • placed in mold, filled with melted paraffin; after final bath.
  • placed in cold water bath for rapid hardening.
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9
Q

How would you prepare thin slice of tissue?

A
  • sectioning: fixed rotary microtome makes slices in fixed distances
  • sharp razor and tubular holder will produce similar results
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10
Q

Why must we stain tissue?

A

-generally tissues are colorless

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11
Q

Steps involved with staining.

A
  • remove paraffin from the slide mounted section with xylene.
  • remove xylene with graded [alcohol] down to water.
  • apply stain.
  • dehydrate with series of alcohol
  • remove alcohol with xylene
  • cement slide together.
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12
Q

Hematoxylin and eosin

A
  • used to display structural features.

- not much about chemical characteristics of the tissue.

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13
Q

Eosin

A
  • stains cytoplasmic components and extracellular

- yellow to pinkish

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14
Q

What do orcein and resorcin fuchsin stains reveal?

A

-elastic material

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15
Q

Silver Impregnation

A

-show reticular fibers and basement membranes

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16
Q

Lipids

A
  • sudan will show the lipid.

- preservation requires technique without use of alcohol (dehydration)

17
Q

How do basic dyes cause staining?

A
  • react with anionic groups in tissue.
  • phosphate, sulfate, carboxyl.
  • higher pH allows more availability for binding.
18
Q

List examples of basic dyes.

A
  • methyl green
  • methylene blue
  • pyronine G
  • toluidine blue
19
Q

How do acid dyes bind?

A
  • form electrostatic linkages with cationic groups. amino groups
  • use in sequence to provide different results.
20
Q

Acid dye examples.

A
  • acid fuchsin
  • aniline blue
  • eosin
  • orange g
21
Q

Metachromasia

A

-dye changes color after reacting with tissue

22
Q

What are histochemical staining techniques used for?

A

-study chemistry of cells and tissues

23
Q

Schiff reagent reaction

A

-reacts with aldehyde groups to form a deep-pink color after exposure to HCl

24
Q

Periodic acid-Schiff reaction

A

-periodic acid cleaves bonds of carbohydrates to form aldehyde group

25
Q

How can RNA organelles be stained?

A
  • use of basic dyes

- require control slides to note basophilic substances

26
Q

Immunocytochemical techniques are used for?

A

-detect the presence of antigens by monoclonal antibodies

27
Q

Antigens

A
  • proteins
  • glycoproteins
  • proteoglycans
28
Q

Antibodies

A

-activated B cells form monoclonal antibodies ( specific)

29
Q

How are antibodies stained?

A
  • fluorescent dye
  • conjugated with visible substance for light microscopy
  • gold or ferritin to produce visible marker in TEM
30
Q

What is indirect labeling?

A

-marker attached to second antibody, specific to antibody which located the antigen of interest.

31
Q

Steps taken in order to visualize tissue on a slide.

A
  • fixing
  • dehydration
  • alcohol removal
  • embedding
32
Q

What is the process of staining?

A

1) remove paraffin with xylene.
2) remove xylene with gradient [alcohol]
3) apply stain and dehydrate with [alcohol]
4) clear alcohol with xylene
5) cement slide to prevent changes

33
Q

What is a hybridoma?

A

-Fusion of activated B cell and myeloma.

34
Q

Acid Fixative

A

-fix chromatin, nucleoli, spindle fibers

35
Q

Carnoy’s Fluid

A

Acid fixative. alcohol, chloroform, glacial acetic acid. Preserves glycogen.

36
Q

Zenker’s Fluid

A

Acidic. potassium dichromate, mercuric chloride, glacial acetic acid. Used for high detail.

37
Q

Bouin’s Fluid

A

picric acid, formalin, glacial aceitc acid. Prolonged washing process.

38
Q

Zirkle-Erliki Fixative

A

potassium dichromate, ammonium dichromate, copper sulfate, distilled water.

Histology (19 decks)

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