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CC and HISTO PLEASE LAST NA > HISTO RA > Flashcards

HISTO RA Flashcards

1
Q

It is a step in tissue processing that involves preservation of tissues as close to original as possible

A

FIXATION

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2
Q

Primary Goal of Fixation

A

to preserve cells and tissues as close as to the original as possible

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3
Q

Secondary Goals of Fixation (2)

A
  1. to HARDEN tissues to facilitate EASY cutting
  2. to PROTECT the tissue from the TRAUMA of further HANDLING
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4
Q

FACTORS THAT INFLUENCE FIXATION

A
  1. Thickness/ Size
  2. Hydrogen Ion Concentration
  3. Osmolality
  4. Concentration of Fixative
  5. Volume
  6. Time and Duration
  7. Temperature
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5
Q

THICKNESS of tissue in FIXATION

A

3-5 mm

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6
Q

SIZE of tissue in FIXATION and also in LUNG specimen

A

3x2 cm; 1-2 cm

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7
Q

Hydrogen Ion Concentration of tissue in FIXATION

A

Ideal pH between 6-8

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8
Q

IDEAL Osmolality of tissue in FIXATION

A

SLIGHTLY HYPERTONIC

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9
Q

In OSMOLALITY, HYPERTONIC solution causes

A

CELL SHRINKAGE

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10
Q

In OSMOLALITY, HYPOTONIC solution causes

A

CELL SWELLING

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11
Q

BUT in PRACTICE osmolality can be

A

ISOTONIC SOLUTION

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12
Q

T/F
In Concentration of Fixative, We use concentrated solution as it can be damaging to tissues

A

FALSE; WE NEVER USE

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13
Q

In Concentration of Fixative, this is diluted to produce 10% formalin

A

FORMALDEHYDE

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14
Q

In Concentration of Fixative, GLUTARALDEHYDE is diluted to produce and is used for

A

a. 3% Glutaraldehyde – Electron microscopy
b. 0.25% – Immunoelectron microscopy

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15
Q

The maximum effective or VOLUME of tissue

A

20 TIMES the volume of specimen (routine)

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16
Q

Fixative to Tissue Ratio:

A

15-20:1

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17
Q

Penetration Rate of Formalin:

A

1 mm/ hour

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18
Q

Volume of Specimen for Osmium Tetroxide (Electron Microscopy) and reason for this

A

5-10 times the volume of specimen as it is quite expensive

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19
Q

For Museum Preparations:

A

should not be less than 50-100 times the volume of the specimen

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20
Q

Time and Duration of tissues

A

Usually subjected to fixative within 24-48 hours

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21
Q

Temperature of tissue for MANUAL FIXATION

A

ROOM TEMPERATURE

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22
Q

FIXATION Temperature of tissue for EM and Histochemistry

A

0-4 degC

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23
Q

FIXATION Temperature of tissue for AUTOTECHNICON and why?

A

40 degC (hastens fixation)

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24
Q

T/F
Does application of heat shortens fixation time

A

TRUE

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25
Q

Temperature range that will ACCELERATE fixation:

A

37-56 degC

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26
Q

FACTORS TO CONSIDER WHEN CHOOSING THE RIGHT FIXATIVE

A
  1. Urgency of the case/ need for immediate examination
  2. Type of specimen/ tissue to be processed
  3. Tissue structure to be studied
  4. Type of section to be made
  5. Staining technique to be applied
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27
Q

In Urgency of the case/ need for immediate examination, you must use a fixative that is ____

A

FAST-ACTING

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28
Q

In Urgency of the case/ need for immediate examination, this is used for rapid in action, thus it’s the most commonly used fixative

A

FORMALIN

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29
Q

Type of specimen/ tissue to be processed that is for preservation of brain tissue for rabies

A

Carnoy’s & Acetone

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30
Q

Type of specimen/ tissue to be processed that is not used for kidneys

A

Bouin’s

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31
Q

In Tissue structure to be studied, it is for glycogen

A

Picric acid fixatives (Brasil’s fluid)

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32
Q

In Tissue structure to be studied, for preservation of fats it is not used as it dissolve fats

A

ACETONE

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33
Q

In Tissue structure to be studied, it is for enzymes, lipases, phosphatase

A

ACETONE

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34
Q

Types of section to be made

A
  1. Serial
  2. Individual
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35
Q

Staining technique to be applied that is a ROUTINE STAIN

A

H&E

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36
Q

This can inhibit HEMATOXYLIN

A

OSMIUM TETROXIDE

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37
Q

FACTORS THAT WILL RETARD OR PROLONG FIXATION PROCESS

A
  1. Cold Temperature
  2. Presence of blood and mucus
  3. Presence of Fat/ Fatty Tissues
  4. Size and thickness
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38
Q

FILL IN THE BLANKS (RETARD OR PROLONG FIXATION)
1. Cold Temperature: it can _____ enzymes
2. Presence of blood and mucus: Remedy ___
3. Presence of Fat/ Fatty Tissues: Remedy ___
4. Size if it is ___ and thickness if it is ___

A

can inactivate enzymes
can be removed by washing the tissue with NSS
cut the specimen thinly to ensure rapid entry of fixative
larger; thicker

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39
Q

FACTORS THAT WILL ACCELERATE OR SHORTENS FIXATION PROCESS

A
  1. Heat Application
  2. Agitation
  3. Size & Thickness
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40
Q

FILL IN THE BLANKS (ACCELERATE OR SHORTENS FIXATION)
1. Heat Application ____; beyond this temp, it can lead to _____
2. Agitation
3. Size if it is _____ & Thickness if it is ___

A

37-56°C ONLY; damage
continuous mixing (autotechnicon)
smaller; thinner

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41
Q

Problem in FIXATION:
Loss of substances soluble in fixing agent/ Loss or inactivation of enzyme; Cause

A

May result from wrong choice of fixative

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42
Q

Problem in FIXATION:
Soft tissues & feather like consistency; Cause

A

Incomplete washing of fixative

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43
Q

Problem in FIXATION:
Brittle & hard tissues; cell shrinkage & swelling; Cause

A

Overfixation/ prolonged fixation

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44
Q

Problem in FIXATION:
Presence of artifacts on tissues; Cause

A

Inadequate or incomplete washing

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45
Q

Problem in FIXATION:
Failure to arrest early autolysis; Cause

A

Insufficient fixative; failure to fix abruptly

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46
Q

FIXATION METHODS

A
  1. Physical Method
  2. Chemical Method
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47
Q

TYPES of PHYSICAL METHOD

A

HEAT FIXATION
MICROWAVE TECHNIQUE

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48
Q

Type of Physical Method that involves thermal coagulation of proteins

A

HEAT FIXATION

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49
Q

FIXATION METHOD that is commonly done and uses chemical fixatives

A

CHEMICAL METHOD

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50
Q

TYPE OF FIXATIVES

A

I. According to Mechanism of Action (as to manner of preservation)
II. According to Action (as to purpose)
II. According to Active Component Substance (as the main ingredient)

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51
Q

FILL IN THE BLANKS
I. According to Mechanism of Action: as to ____
II. According to Action: as to ____
II. According to Active Component Substance: as the ____

A

manner of preservation
purpose
main ingredient

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52
Q

Types of FIXATIVES According to Mechanism of Action (as to manner of preservation)

A

ADDITIVE FIXATIVES
NON-ADDITIVE FIXATIVES

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53
Q

Type of FIXATIVES According to Mechanism of Action (as to manner of preservation) that when used are combined or absorbed by the tissues

A

ADDITIVE FIXATIVE

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54
Q

In ADDITIVE FIXATIVES when used are combined or absorbed by the tissues it becomes ____

A

part of the tissue

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55
Q

ALL the fixatives available are additive except:

A

Acetone & Alcohol

56
Q

Type of FIXATIVES According to Mechanism of Action (as to manner of preservation) that do not bind with the tissue

A

NON-ADDITIVE FIXATIVES

57
Q

In NON-ADDITVE FIXATIVES it does not bind with the tissue for it is ____

A

not absorbed or taken in by the tissue

58
Q

Examples of NON-ADDITIVE FIXATIVES

A

Acetone and Alcohol

59
Q

Types of FIXATIVES according to Action (as to purpose)

A
  1. Microanatomical
  2. Cytological
  3. Histochemical
60
Q

Type of FIXATIVES according to Action (as to purpose) that allows general microscopic study of tissue structure

A

MICROANATOMICAL

61
Q

Type of FIXATIVES according to Action (as to purpose) that preserve parts of the cell (either nucleus or cytoplasm)

A

CYTOLOGICAL

62
Q

Types of FIXATIVES in Cytological

A
  1. Nuclear
  2. Cytoplasmic
63
Q

Fixative that is for nuclear chromatin, chromosomes, etc.

A

NUCLEAR FIXATIVE

64
Q

NUCLEAR FIXATIVE
___ glacial acetic acid
pH with ___ 4.6

A

WITH
LESS THAN

65
Q

Nuclear Fixative EXAMPLES

A

Bouin’s
Carnoy’s
Heidenhein’s Susa
Newcomer’s
Flemming’s with HAc

66
Q

Fixative that is for golgi body, mitochondria, etc.

A

CYTOPLASMIC FIXATIVES

67
Q

CYTOPLASMIC FIXATIVE
___ glacial acetic acid
pH with ___ 4.6

A

WITHOUT
MORE THAN

68
Q

Cytoplasmic Fixatives EXAMPLES

A

Regaud’s/ Moller’s
Orth’s
Formalin with post chroming
Flemming’s without HAc

69
Q

Type of FIXATIVES according to Action (as to purpose) that preserve the chemical components of tissues like enzyme

A

HISTOCHEMICAL

70
Q

EXAMPLES of Histochemical

A

Acetone
Absolute ethyl alcohol
Newcomer’s
10% Formol Saline

71
Q

Types of Fixatives According to Active Component Substance (as the main ingredient)

A
  1. Aldehyde Fixatives
  2. Metallic Fixatives
  3. Picric Acid Fixatives
  4. Alcoholic Fixatives
  5. Glacial Acetic Acid
  6. Acetone
  7. Osmium Tetroxide
  8. Trichloacetic Acid
72
Q

Types of Aldehyde Fixatives

A

A. FORMALDEHYDE/ FORMALIN FIXATIVES
B: GLUTARALDEHYDE FIXATIVES
C: GLYOXAL

73
Q

Types of FORMALDEHYDE/ FORMALIN FIXATIVES

A

a. 10% Formalin
b. 10% Formol Saline
c. 10% Neutral Buffered Formalin
d. Formol Corrosive/ Formol Sublimate
e. Alcoholic Formalin/ Gendres

74
Q

ROUTINELY USED Formaldehyde/ Formalin Fixative

A

10% Formalin

75
Q

10% Formalin preparation:

A

1 L or 1000 mL Formalin:
Combine 100 ml of 37-40% formalin + 900 ml distilled water

76
Q

Formaldehyde/ Formalin Fixative that is to preserve CNS and post mortem tissues

A

10% Formol Saline

77
Q

10% Formol Saline uses ____ as diluent

A

NSS

78
Q

Formaldehyde/ Formalin Fixative that is for tissues with iron pigments

A

10% Neutral Buffered Formalin

79
Q

10% Neutral Buffered Formalin preparation

A

Same with 10% formalin, but sodium phosphate was added as buffer

80
Q

Formaldehyde/ Formalin Fixative that is best for lipids & neutral phospholipids

A

Formol Corrosive/ Formol Sublimate

81
Q

Formol Corrosive/ Formol Sublimate preparation

A

Formaldehyde + Mercuric Chloride

82
Q

Formaldehyde/ Formalin Fixative that is for sputum and microincineration techniques

A

Alcoholic Formalin/ Gendres

83
Q

Alcoholic Formalin/ Gendres preparation

A

Combination of 95% Ethanol + Glacial Acetic Acid (HAc) + Formaldehyde

84
Q

Disadvantage of Formalin:

A

It may precipitate upon longer storage

85
Q

To prevent precipitation of formaldehyde to white paraformaldehyde:

A

Add 10% Methanol

86
Q

Aldehyde Fixative that is for Electron Microscopy

A

GLUTARALDEHYDE FIXATIVE

87
Q

Types of GLUTARALDEHYDE FIXATIVES

A

a. 2-5% Solution
b. 4% Solution

88
Q

Glutaraldehyde fixative that is for small tissue fragments/ needle biopsies

A

2-5% Solution

89
Q

Glutaraldehyde fixative that is for larger tissues less than 4 mm thick

A

4% Solution

90
Q

Aldehyde Fixative that is SMALLEST ALDEHYDE

A

GLYOXAL

91
Q

Types of Metallic fixatives

A

A. MERCURIC CHLORIDE
B. LEAD FIXATIVES
C. CHROMATE FIXATIVES

92
Q

It is the most common metallic fixative

A

MERCURIC CHLORIDE

93
Q

Metallic Fixative that is excellent for trichrome staining & tissue photography

A

MERCURIC CHLORIDE

94
Q

Types of MERCURIC CHLORIDE

A

a. Zenker’s Fluid
b. B5
c. Heidenhein’s Susa
d. Zenker Formol/ Helly’s Fluid

95
Q

MERCURIC CHLORIDE fixative that is for Liver, spleen, connective tissue fibers & nuclei

A

Zenker’s Fluid

96
Q

MERCURIC CHLORIDE fixative that is for bone marrow

A

B5

97
Q

MERCURIC CHLORIDE fixative that is for Tumor skin biopsies and an excellent cytologic fixative

A

Heidenhein’s Susa

98
Q

MERCURIC CHLORIDE fixative that is for Pituitary gland, bone marrow, spleen, and liver

A

Zenker Formol/ Helly’s Fluid

99
Q

DISADVANTAGE of MERCURIC CHLORIDE:

A

They produce black mercury deposits

100
Q

REMEDY for MERCURIC CHLORIDE:

A

Alcohol Iodine is used to wash out excess mercuric fixatives

101
Q

It is proposed as mercuric chloride substitute as it is toxic to man

A

ZINC SULFATE

102
Q

METALLIC FIXATIVE that is for acid mucopolysaccharide

A

LEAD FIXATIVE

103
Q

METALLIC FIXATIVE that have potassium
dichromate

A

CHROMATE FIXATIVES

104
Q

Types of CHROMATE FIXATIVES

A

a. 1-2% Chromic Acid
b. 3% Potassium Dichromate
c. Regaud’s/ Moller’s
d. Orth’s Fluid

105
Q

CHROMATE FIXATIVE that is for carbohydrates

A

1-2% Chromic Acid

106
Q

CHROMATE FIXATIVE that is for lipids &
mitochondria

A

3% Potassium Dichromate

107
Q

CHROMATE FIXATIVE that is for mitochondria, RBC & colloid containing tissues

A

Regaud’s/ Moller’s

108
Q

CHROMATE FIXATIVE that is for Rickettsiae & other bacteria, tissue necrosis

A

Orth’s Fluid

109
Q

Type of Fixative According to Active Component Substance (as the main ingredient) that is an excellent for glycogen demonstration

A

PICRIC ACID FIXATIVES

110
Q

This is used as fixative, decalcifying agent, and stain

A

PICRIC ACID FIXATIVES

111
Q

Types of PICRIC ACID FIXATIVES

A

a. Bouin’s Solution
b. Brasil’s Alcoholic Picformol
c. Hollande’s Solution

112
Q

Type of Picric Acid Fixative that is for embryos, pituitary biopsies & endometrial curetting

A

Bouin’s Solution

113
Q

T/F
Bouin’s Solution is for KIDNEY STRUCTURES

A

FALSE; IT IS NOT

114
Q

T/F
Bouin’s Solution is compatible with feulgen’s stain

A

FALSE; IT IS NOT

115
Q

This is used to wash out excess amount of picric acid fixatives

A

50-70% Alcohol

116
Q

This acts as both fixative and dehydrating agent

A

ALCOHOL

117
Q

Types of ALCOHOL FIXATIVES

A

a. 95% Isopropyl Alcohol/ IPA
b. Methyl Alcohol (100%)
c. Ethyl Alcohol (70-100%)
d. Carnoy’s
e. Newcomer’s

118
Q

Type of ALCOHOL FIXATIVE that is for touch preparations

A

95% Isopropyl Alcohol/ IPA

119
Q

Type of ALCOHOL FIXATIVE that is the MOST RAPID tissue fixative

A

CARNOY’S

120
Q

Type of ALCOHOL FIXATIVE that is for fixing chromosomes & lymph glands

A

CARNOY’S

121
Q

Type of ALCOHOL FIXATIVE that is for mucopolysaccharides (histochemical) & nuclear protein

A

NEWCOMER’S

122
Q

Type of Fixative According to Active Component Substance (as the main ingredient) that solidifies at 17 degC

A

GLACIAL ACETIC ACID

123
Q

It is Contraindicated for cytoplasmic fixation as it is a fixative for nucleus

A

GLACIAL ACETIC ACID

124
Q

This is used in combination with other fixatives to produce a compound fixative (but can be used alone)

A

GLACIAL ACETIC ACID

125
Q

Type of Fixative According to Active Component Substance (as the main ingredient) that is used both fixative & dehydrating agent

A

ACETONE

126
Q

This must be used at ICE COLD TEMPERATURE between -5 to 4 degC

A

ACETONE

127
Q

This is for phosphatases & lipases enzymes

A

ACETONE

128
Q

DISADVANTAGE of ACETONE

A

Evaporates easily & dissolves fats

129
Q

Type of Fixative According to Active Component Substance (as the main ingredient) that can be used for myelin & peripheral nerves

A

OSMIUM TETROXIDE

130
Q

Type of Fixative According to Active Component Substance (as the main ingredient) that is a fixative & a weak and slow decalcifying agent

A

TRICHLOROACETIC ACID

131
Q

Fixatives for brain tissues for the diagnosis of rabies

A
  1. Carnoy’s Fluid – MOST RAPID FIXATIVE
  2. Acetone
132
Q

Fixatives for Enzyme Histochemistry

A
  1. 4% Formaldehyde
  2. Formol Saline
133
Q

Fixatives for Electron Microscopy

A
  1. Osmium Tetroxide
  2. Glutaraldehyde
  3. Paraformaldehyde
134
Q

Fixatives for Enzyme Histochemistry & Electron Immunocytochemistry

A
  1. Karnovsky’s
  2. Glutaraldehyde
  3. Paraformaldehyde
135
Q

Fixatives often used in conjunction with other fixatives to produce a compound fixative

A
  1. Glacial acetic acid
  2. Trichloroacetic acid/ TCA

CC and HISTO PLEASE LAST NA (4 decks)

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