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Cell Bio > Med Application of Tissue Processing > Flashcards

Med Application of Tissue Processing Flashcards

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1
Q

What are all the steps in tissue processing?

A

1) Fixation
2) Dehydration
3) Clearing
4) Embedding
5) Sectioning
6) Mounting
7) Staining

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2
Q

What chemical is used in the fixation step of tissue processing?

A

Formalin

Glutaraldehyde (complex aldehyde)

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3
Q

What is the purpose of the fixation step of tissue processing?

A

To preserve normal tissue architecture after death. Cross-links the proteins and stabilizes them.

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4
Q

What chemical is used in the dehydration step of tissue processing?

A

Graded series of ethanol

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5
Q

What is the purpose of the dehydration step of tissue processing?

A

To remove water from the tissue specimen and replace it with alcohol

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6
Q

What chemical is used in the clearing step of tissue processing?

A

Xylene

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7
Q

What is the purpose of the clearing step of tissue processing?

A

Prepares the tissue for the embedding medium (e.g. paraffin)

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8
Q

What chemical is used in the embedding step of tissue processing?

A

Paraffin

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9
Q

What is the purpose of the embedding step in tissue processing?

A

The tissue is infiltrated with paraffin so that it is hard enough to section

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10
Q

What is the purpose of the sectioning step in tissue processing?

A

The tissue is cut into thin sections with a microtome

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11
Q

What is the purpose in the mounting step of tissue processing?

A

The cut paraffin section is placed on a glass microscope slide

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12
Q

What is the purpose in the staining step of tissue processing?

A

To impart contrast to tissue structures so that they may be distinguished upon microscopic observation

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13
Q

What are acidophilic structures?

A

Structures or tissues that have an affinity for negative charge dye.

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14
Q

What are basophilic structures?

A

Structures or tissues that have an affinity for positive charge dye.

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15
Q

What is H&E?

A

Hematoxylin and eosin

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16
Q

What structures does the H in H&E dye and in what color?

A

(Hematoxylin) RNA, DNA, ribosomes, and rER

Blue

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17
Q

What structures does the E in H&E dye and in what color?

A

(Eosin) Secretory vesicles, sER, lysosomes, mitochondria, and type I collagen
Pink

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18
Q

In Feulgen reaction, what structure does it dye and in what color?

A

DNA

Magenta

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19
Q

In Mallory triple (trichrome), what structures does it have affinity for?

A

nuclei, muscle, collagen, hyaline cartilage

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20
Q

In Mallory triple, what color will be imparted on the nuclei?

A

red

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21
Q

In Mallory triple, what color will be imparted on muscle?

A

red to orange

22
Q

In Mallory triple, what color will be imparted on collagen?

A

blue

23
Q

In Mallory triple, what color will be imparted on hyaline cartilage?

A

blue

24
Q

In PAS reaction, what structure does it have affinity for and what color will be imparted?

A

Carbohydrates. Magenta

25
Q

In Osmic acid, what structure does it have affinity for and what color will be imparted?

A

lipids, black

26
Q

In Verhoeff stain, what structure will it have affinity for and what color will be imparted?

A

Elastic fibers, black

27
Q

In silver methods, what structures will it have affinity for and what color will be imparted?

A

intermediate filaments of nerve cells, glial cells and reticular fibers. Black

28
Q

In trypan blue, what structure does it have affinity for and in what color?

A

macrophages, used on live tissues, blue

29
Q

In Prussian blue stain, what structure does it stain and in what color?

A

Hemosiderin (ferric iron), blue

30
Q

In Nissl stain, what structure does it stain and in what color?

A

ribosomes, blue

31
Q

In iron hematoxylin stain, what structures are stained and in what color?

A

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

32
Q

What stain should you use for coagulative necrosis?

A

H & E

33
Q

What stain should be used to detect thickened basement membranes in kidney disease?

A

PAS

34
Q

What stain should be used to highlight fibrosis?

A

mallory triple

35
Q

What stain can be used to demonstrate nuclear changes in cancer?

A

feuglen reaction stain

36
Q

What stain can be used to demonstrate excessive iron accumulation in hemochromatosis?

A

prussian blue

37
Q

What stain can be used to identify extracellular deposits of amyloid?

A

congo red

38
Q

What stain can be used to detect elastic fibers often found in Marfan syndrome?

A

Verhoeff

39
Q

What is metachromasia?

A

It is a phenomenon in which a given stain imparts different colors to the tissue.

40
Q

What is immunocytochemistry (immunohistochemistry)?

A

Technique that uses an antibody tagged with a marker directed to a specific antigen (e.g. fluorescent molecule tagged to a protein associated with the cell or tissue)

41
Q

What is the direct method of immunocytochemistry?

A
  • Detects presence of antigen, regardless of antigen’s location on cell surface or within nucleus, using an antibody tagged with a marker and specific to the antigen
  • Less sensitive than indirect method as fewer antibodies bind to the antigens
42
Q

What is the indirect method of immunocytochemistry?

A
  • Uses primary and secondary antibodies. The first one is not tagged and is specific to the antigen of interest. The second is tagged and is specific to the primary antibody
  • More sensitive as several labeled anti-antibodies can bind to the primary antibody, thus amplifying the signal
43
Q

What is the medical application of immunocytochemistry?

A

Used to categorize tumor origins and tumor aggressiveness

44
Q

What is in situ hybridization?

A

Identifies a specific genetic sequence

45
Q

What is the medical application of in situ hybridization?

A

Used to identify a cell infected with a virus or specific genes on a chromosome. Can also look for extra copies of genes (amplification).

46
Q

When are frozen tissue sections used?

A

when urgent tissue diagnosis is needed

47
Q

What procedure can be used to categorize tumor origin and tumor aggressiveness?

A

immunocytochemistry

48
Q

What technique can be used to detect HER2/neu receptors?

A

immunocytochemistry

49
Q

What is the goal of in situ hybridization?

A

to determine whether a known specific genetic sequence is present

50
Q

What stain should be used to detect glycogen storage disease in alpha1-antitrypsin deficiency in liver cells?

A

PAS

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