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MTAP 2 - HISTOPATH > Fixation > Flashcards

Fixation Flashcards

1
Q

Most critical step in preserving tissue

A

Fixation

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2
Q

initial step in tissue processing

A

Fixation

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3
Q

Purpose of fixation

A

Killing, penetration, and Hardening

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4
Q

Primary goal of Fixation

A

Preserve tissue and cells

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5
Q

Secondary Goal of fixation

A
  1. Hardens tissue
  2. Protect tissue from further trauma
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6
Q

Physical Method of Fixation

A

Heat Fixation
Microwave Fixation

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7
Q

Heat fixation is usually used in

A

Microbiological Section (Bacterial smear)
For FROZEN section only

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8
Q

Neurochemical substance preserve by Microwave technique

A

Acetylcholine

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9
Q

Use of fixative agent is under what method

A

Chemical Method

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10
Q

Fixation is usually carried out at what temp

A

Room temperature

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11
Q

Formalin temp. Used for urgent biopsies

A

Formalin @60

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12
Q

Formalin @100 deg. Cel. Is required for

A

Fixing tissues with TUBERCULOSIS

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13
Q

Required size for tissue

A

3x2 cm

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14
Q

Required thickness for tissue

A

3-5mm

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15
Q

Common osmolality requirement used in Lab

A

Isotonic

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16
Q

Concentration used in Formalin

A

10% Formalin

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17
Q

Ideal time to perform fixation

A

20-30 mins following interruption of blood supply

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18
Q

Required osmolality in fixation based on book

A

Slightly Hypertonic

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19
Q

Concentration used in Formalin

A

10%

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20
Q

Measurements to form 10% formalin

A

100 ml 37%-40% formalin + 900 mL distilled water

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21
Q

Disdvantage of formalin

A

Easy to precipitate

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22
Q

Substance added to avoid precipitation of formalin

A

Add 10% methanol

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23
Q

Concentration of glutaraldehyde for EM

A

3%

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24
Q

Causes for retarded fixation

A

Blood
Mucus
Fats
Large tissues

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25
Q

Causes for accelerated fixation

A

Heat (37-56)
Agitation

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26
Q

Common volume ratio of fixative to tissue

A

20:1

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27
Q

Osmium tetroxide volume

A

5-10x:1

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28
Q

Hallow organ must be packed with _____

A

Cotton

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29
Q

Human brain must undergo

A

Intravascular perfusion

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30
Q

Washing out of blood from Human brain uses what fixative

A

Ringers lactate

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31
Q

For large organ such as Uterine, cervix, fibroid it must undergo

A

LENDRUM’S METHOD

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32
Q

Lundrum’s method fixative contains

A

4% aqueous phenol (1-3 days)

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33
Q

Effect of fixative in tissues

A

Harden tissues
Make cell resistant to damage
Act as accentuators and mordants

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34
Q

Incomplete washing may cause

A

Precipitation of artefacts

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35
Q

Too hard and brittle tissues is due to

A

Overfixation

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36
Q

Too soft tissue may be due to

A

Underfixation

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37
Q

Wrong choice of fixative may lead to

A

Loss of soluble substance in fixing agent

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38
Q

Factos needed to be considered when choosing fixatives

A

Needed for an immediate exam
Type of specimen
Structure to be studied
Staining technique
Type of section to be made

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39
Q

Fixative used in brain tissues

A

Carnoys
Acetone

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40
Q

fixative that cannot be used in kidney/spleen

A

Bouins

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41
Q

Fixative used for lipases/enxymes

A

Acetone

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42
Q

Used to preserve glycogen

A

Brasil’s

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43
Q

Mechanism of action:

Type of fixative that is being absorbed by the tissues

A

Additive Fixatives

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44
Q

Mechanism of Action:

Fixatives that are not being absorbed by the tissues

A

Non-Additive Fixatives

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45
Q

Non-additive fixatives are

A

Acetone
Alcohol

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46
Q

Fixative use for general microscopic study

A

MICROANATOMICAL

10% formol saline
10% neutral buffered formalin
Heidanhain’s susa
Formol sublimate/formol corrosive
Zenker’s
Bouin’s
Brasil’s

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47
Q

Nuclear fixative must contain

A

Glacial acetic acid

48
Q

Nuclear fixative pH requirement

A

Less than 4.6 (acidic)

49
Q

Cytoplasmic pH requirement

A

Not more than 4.6

50
Q

Nuclear fixatives are

A

Flemmings with Glacial Acetic Acid
Carnoys
Bouin’s
Newcomer’s
Heidenhain’s Susa

51
Q

Cytoplasmic fixatives are

A

Flemming w/o HAc
Helly’s
Formalin with post chroming
Regaud’s/Moller’s
Orth’s

52
Q

Fixative used to preserve chemical components are called as

A

HISTOCHEMICAL FIXATIVES

53
Q

Histochemical Fixatives are

A

NAFA

Newcomer’s
Absolute Ethyl alcohol
10% Formol saline
Acetone

54
Q

A nuclear fixative, considered as Compound fixative that solidifies at 17 deg. Cel.

A

Glacial Acetic acid

55
Q

What type of fixative is trichroloacetic acid

A

Compound fixative

56
Q

Tricholoroacetic acid is both used as

A

Fixative and Decalcifying agent

57
Q

Fixative used at ice cold temperature, ideal for enzymes (lipase, phosphatase)

A

Acetone

58
Q

Acetone is both used as

A

Fixative and dehydrating agent

59
Q

Ideal for enzymes and diagnosis of rabies

A

Acetone

60
Q

Disadvantages of Acetone are

A

Dissolves fats
Evaporates easily
Raw material for shabu

61
Q

Fixative that is expensive and NOT FAST ACTING used in EM

A

Osmium tetroxide

62
Q

Osmium tetroxide preserves

A

Myelin and Peripheral nerves

63
Q

Fixatives with Osmium Tetroxide are

A

Flemmings with HAc (nuclear)
Flemmings w/o HAc (cytoplasm)

64
Q

Used as Fixative, Stain, DECALCIFYING

A

Picric Acid

65
Q

Excellent for Glycogen Demonstration

A

Picric Acid Fixative

66
Q

Disadvantage of Picric Acid

A

Produced yellow color when used as fixative

67
Q

Remedy for yellow stain while using picric acid

A

70% alcohol + LITHIUM CARBONATE — washed w/water

70% ethanol + sodium thiosulfate (washed w/ water

68
Q

Fixatives with Picric Acid are

A

Bouin’s solution
Brasil’s Alcoholic formalin
Hollande’s solution

69
Q

Fixative used in embryo and pituitary biopsies, and endometrial curettings

A

Bouin’s solution

70
Q

Reason as to why Bouin’s solution is not used in kidneys

A

Inhibit Feulgen Reaction (dmeonstrate RNA, DNA)

71
Q

Hollande’s solution is used for

A

GIT biopsies & endocrine tissues

72
Q

Fixative that easily disintegrate and precipitates protein

A

Alcohol Fixative

73
Q

Alcohol is used both as

A

Fixative, dehydrating

74
Q

Alcohol is ideal for

A

Small tissue fragments

75
Q

Disadvantage of Alcohol

A

Glycogen polymerization

76
Q

Alcohol fixative used for wet and dry smear, BM tissues

A

Methyl alcohol (100%)

77
Q

Isopropyl alcohol is used for

A

Touch preparation

78
Q

Ethyl alcohol 50-70% is used for

A

Blood, tissue film and smear

79
Q

Fixative used for chromosomes and lymph glands; brain tissue

A

Carnoy’s

80
Q

Considered as a fast-acting fixative

A

Carnoy’s

81
Q

Used for histochemical and nuclear fixative

A

Newcomer’s

82
Q

Used for CT mucin and Umbilical cord

A

Rossman’s solution

83
Q

Most common metallic fixative

A

Mercuric chloride

84
Q

Ideal staining for mercuric chloride

A

Trichrome staining

85
Q

Aids in black mercury deposit from mercuric chloride

A

Alcoholic iodine

86
Q

Component of Zenker’s fluid

A

Mercuric chloride, Glacial acetic acid

87
Q

Structures preserved by Zenker’s fluid

A

Liver/spleen, Connective tissues fibers, nuclei

88
Q

Zenker’s formol is also known as

A

Helly’s fluid

89
Q

Structures preserved by Zenker’s formol

A

Pituitary gland, BM, other organ with blood supply

90
Q

Component of Zenkers Formol

A

Potassium dichromate
40% formaldehyde

91
Q

Used for tumor biopsies

A

Heidenhains susa

92
Q

Component of Heidenhains susa

A

TCA, HAc, formalin

93
Q

With anhydrous sodium acetate

A

B5

94
Q

Tissue preserved by B5

A

Bone marrow

95
Q

Preserves carbohydrates

A

Chromic acid (1-2%)

96
Q

Potassium dichromate preserves

A

Lipids and mitochondria

97
Q

Other name for Regaud’s

A

Moller’s

98
Q

Regauds fixed what structure

A

Chromatin, golgi bodies, mitotic figures, mitochondria,RBC containing colloid tissues

99
Q

Fixative that preserves ricketssial tissue, necrosis

A

Orth’s

100
Q

Fixative used for tissue mucin

A

Lead fixative

101
Q

Fixative for acid mucopolysaccharide

A

Lead fixative

102
Q

Routinely used fixative

A

10% formalin

103
Q

Component of 10% formol saline

A

Sodium chloride

104
Q

Structure preserve by 10% formol saline

A
  • Histochemical
  • General post mortem tissue
  • CNS tissue
105
Q

Composed with phosphate buffer + 10% formalin

A

10% neutral buffered saline

106
Q

Structure preserved by 10% neutral buffered saline

A

Iron pigment
Elastic fiber

107
Q

Component Of formol corrosive

A

Mercuric chloride + formaldehyde

108
Q

Structure preserve by Formol corrosive

A

Lipids
Phospholipids
Neutral fats

109
Q

Other term for Alcoholic Formalin

A

Gendre’s solution

110
Q

Component of Alcoholic formalin

A

FO-PEG

FOrmadehyde
Picric Acid
95% Ethyl Alcohol
Glutaraldehyde

111
Q

Tissue preserved by alcoholic formalin

A

Sputum specimen

112
Q

Ideal for EM and histochemistry

A

Glutaraldehyde

113
Q

Fixatives used in Enzyme Histochemistry

A

4% formaldehyde
Formol saline

114
Q

Fixatives for EM

A

POG

Paraformaldehyde
Osmium tetroxide
Glutaraldehyde

115
Q

Smallest aldehyde fixative and fast acting

A

Glyoxal

116
Q

Fixative used for Electron Histochemistry and Electron Immunocytometry

A

Karnovsky’s paraformaldehyde-glutaraldehyde

MTAP 2 - HISTOPATH (5 decks)

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