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HISTOPATHOLOGY > SEMIS > Flashcards

SEMIS Flashcards

1
Q

What is the first step in risk management within a hospital facility?

A

Identify all hazards in and emanating from the laboratory

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2
Q

List the top 6 workplace hazards.

A
  • Biological
  • Physical
  • Chemical
  • Psychological
  • Ergonomic
  • Safety
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3
Q

What must standard operating procedures (SOP) include?

A

Control of hazardous substances, risk assessments, and other health and safety information

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4
Q

What is a planar concave microtome knife used for?

A

Cutting very soft samples like fascia, tendons, adipose tissues, and connective tissues

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5
Q

What types of materials are wedge profile knives used for?

A

Moderately hard materials such as epoxy or cryogenic samples

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6
Q

What is the function of a cryostat?

A

To preserve frozen tissue samples and slice them thin enough for microscopic examinations

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7
Q

What is the difference between formaldehyde and formalin?

A

Formaldehyde is a gas that is dissolved in water to form formalin, which is a saturated solution of formaldehyde

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8
Q

Fill in the blank: The __________ is an instrument for cutting extremely thin sections for examination under a microscope.

A

Microtome

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9
Q

What are the major equipment types used in histotechnology?

A
  • Microscope
  • Microtome
  • Cryostat
  • Autotechnicon
  • Automated coverslipper
  • Automated H and E stainer
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10
Q

True or False: Poor equipment handling can affect the diagnosis and prognosis of a patient.

A

True

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11
Q

What should be maintained for every piece of laboratory equipment?

A

A current file containing name, manufacturer, model number, serial number, and maintenance records

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12
Q

What is the hazard associated with acetic acid?

A

Irritation to skin, eyes, and respiratory system upon direct contact

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13
Q

What is the recommended storage condition for isopentane?

A

In a refrigerator or freezer suited for explosive atmospheres

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14
Q

List three types of staining protocols mentioned.

A
  • 70% Ethanol for tissue fixation
  • Haematoxylin for nuclease staining
  • Eosin stain for cytoplasm staining
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15
Q

What is the risk associated with chloroform?

A

Toxic when inhaled or ingested, and can cause disorientation, loss of consciousness, and death

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16
Q

What should be done with concentrated acids?

A

They should be added to water, not water to acid, to prevent splashing

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17
Q

What is the purpose of a smear preparation in cytological exams?

A

To spread cellular materials over a slide for examination

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18
Q

What are the effects of prolonged exposure to toluene?

A

Impaired memory, poor coordination, mood swings, and permanent nerve damage

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19
Q

What is the characteristic of picric acid when dry?

A

It is explosive when dry or combined with metals

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20
Q

Fill in the blank: The method used to immerse a tissue specimen in isotonic saline solution for examination is called __________.

A

Teasing or dissociation

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21
Q

What is the significance of keeping tissue samples intact in histotech?

A

To provide fast and accurate results

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22
Q

What precautions should be taken when handling formaldehyde?

A

Workers should be periodically monitored for exposure levels

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23
Q

What is the purpose of smear preparation?

A

Useful in cytological exams, particularly Pap smears for cancer diagnosis

Smear preparation involves spreading cellular materials lightly over a slide.

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24
Q

Describe the process of smear preparation.

A

Place a drop of secretion on one slide, move two slides in opposite directions to initiate material flow

This is done in one single uninterrupted motion.

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25
Q

What are recommended specimens for smear preparation?

A
  • Serous
  • Concentrated sputum
  • Enzymatic lavage from GIT
  • Blood smears
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26
Q

What is a touch preparation?

A

Tissue is pressed onto a slide to transfer cells directly for examination

Cells can be examined without destroying their intercellular relationships.

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27
Q

What is a frozen section?

A

Fresh tissue frozen immediately for rapid diagnosis

Results are usually obtained within 5-15 minutes.

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28
Q

What types of specimens can be processed as frozen sections?

A
  • Sputum
  • Pleural
  • Peritoneal
  • CSF
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29
Q

What is the purpose of streaking in laboratory procedures?

A

To obtain a uniform distribution of secretion on a slide

Too thick or too thin samples are unsuitable for examination.

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30
Q

Define cryostat.

A

A cold chamber with a built-in microtome that maintains a temperature of -10 to -20˚C.

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31
Q

What are the applications of frozen sections?

A
  • Rapid pathologic diagnosis during surgery
  • Diagnostic and research enzyme histochemistry
  • Demonstration of soluble substances (lipids and carbohydrates)
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32
Q

What is the most rapid method of freezing tissues?

A

Liquid nitrogen

It is used in histochemistry and during operative procedures.

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33
Q

What are some disadvantages of using liquid nitrogen for freezing?

A
  • Soft tissue may crack
  • Overcooling can damage biopsy blocks
  • Uneven cooling can complicate diagnostic interpretation
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34
Q

What is the significance of the fixative in histotechnology?

A

Preserves the morphologic and chemical integrity of cells and prevents decomposition and distortion.

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35
Q

What are the basic mechanisms in fixation?

A
  • Additive fixation
  • Non-additive fixation
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36
Q

What does additive fixation involve?

A

The chemical constituent of the fixative becomes part of the tissue, forming cross-links

Examples include formalin and osmium tetroxide.

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37
Q

What does non-additive fixation involve?

A

The fixing agent alters the tissue composition without becoming part of the tissue

Examples include alcoholic fixatives like ethanol and methanol.

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38
Q

List the main factors in fixation.

A
  • Hydrogen ion concentration
  • Temperature
  • Thickness of section
  • Osmolality
  • Concentration
  • Duration of fixation
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39
Q

What are the effects of fixatives on tissues?

A
  • Harden tissues
  • Make cells resistant to damage
  • Inhibit bacterial decomposition
  • Increase optical differentiation
  • Act as mordants for staining
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40
Q

What are the characteristics of a good fixative?

A
  • Cheap
  • Stable
  • Safe to handle
  • Kill cells quickly
  • Minimal shrinkage
  • Permit rapid penetration
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41
Q

What are the two categories of fixation according to action?

A
  • Microanatomical fixatives
  • Cytological fixatives
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42
Q

What is the purpose of nuclear fixatives?

A

Preserve nuclear structures such as chromosomes

Usually contain glacial acetic acid.

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43
Q

What is the importance of lipid fixation?

A

Lipids are largely removed during tissue preparation; cryostat sections are used for demonstration

Fixatives containing mercuric chloride can effectively preserve lipids.

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44
Q

What fixatives are recommended for glycogen fixation?

A
  • Alcohol-based fixatives
  • Rossman’s fluid
  • Cold absolute alcohol
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45
Q

What are the pros and cons of acrolein as a fixative?

A

Pros: Preserves microanatomy with minimal distortion; Cons: Slow fixative, requires 24 hours or longer.

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46
Q

What is the routine turn-over time for surgical pathology results?

A

24 hours

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47
Q

What is the routine turn-over time for frozen section results?

A

5-15 minutes

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48
Q

What is the recommended storage time for pathology/bone marrow slides?

A

10 years

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49
Q

What must be included in the documentation for pathology reports?

A
  • Surgical Pathology
  • Cytology
  • Autopsy Report
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50
Q

True or False: Fixation is the first and most critical step in histotechnology.

A

True

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51
Q

Fill in the blank: The primary fixation in buffered formalin is usually carried out for _______ hours.

A

2-6

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52
Q

What is Acrolein commonly mixed with for tissue fixation?

A

Glutaraldehyde or formaldehyde

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53
Q

What are the advantages of using Acrolein as a fixative?

A
  • Penetrates and fixes tissues evenly
  • Preserves microanatomic and cytologic details with minimal shrinkage and distortion
  • Preserves enzymes and nucleoproteins
  • Demonstrates fats and mucin
  • Ideal for most staining techniques
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54
Q

What is a major disadvantage of using Acrolein?

A

Slow fixative, requires 24 hours or longer

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55
Q

What is the most widely used fixative in histology?

A

10% formalin

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56
Q

What is the common dilution ratio for preparing 10% formalin?

A

1:10 or 1:20

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57
Q

What is a disadvantage of using unbuffered formaldehyde?

A
  • Reduces basophilic and eosinophilic staining
  • Forms brown pigment granules on blood-containing tissues
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58
Q

What is 10% neutral buffered formalin primarily used for?

A

Preservation and storage of surgical, post-mortem, and research specimens

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59
Q

What are the components of 10% neutral buffered formalin?

A
  • Anhydrous sodium dihydrogen phosphate
  • Anhydrous disodium hydrogen phosphate
  • 40% formaldehyde
  • Distilled water
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60
Q

What is the fixation time for 10% neutral buffered formalin?

A

4-24 hours

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61
Q

What is the purpose of adding sodium chloride to 10% formal saline?

A

To create a simple microanatomical fixative

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62
Q

What are the components of 10% formol saline?

A
  • Saturated formaldehyde (40% by weight volume)
  • Sodium chloride
  • Distilled water
63
Q

What is the recommended fixation time for 10% formol saline?

A

24 hours at 35 °C, 48 hours at 20-25 °C

64
Q

What is a major advantage of formaldehyde as a fixative?

A
  • Cheap
  • Readily available
  • Preserves fat and mucin
65
Q

What is a disadvantage of formaldehyde in tissue fixation?

A

Can cause allergic dermatitis

66
Q

What is the main advantage of glutaraldehyde over formalin?

A

More stable effect on tissues, firmer textures

67
Q

What is the primary use of mercuric chloride in fixation?

A

Commonly used as a secondary fixative

68
Q

What solution is Zenker’s fluid composed of?

A
  • Mercuric chloride stock solution
  • Glacial acetic acid
69
Q

What are the pros of using Zenker’s fluid?

A
  • Fairly rapid and even fixation
  • Compatible with most stains
70
Q

What is a major con of Zenker’s fluid?

A

Poor penetration

71
Q

What does the term ‘inert’ mean in the context of fixatives?

A

Chemically inactive or no chemical reaction

72
Q

What is the purpose of Dezenkerizing?

A

To remove mercuric pigment deposits from tissues

73
Q

List the reagents needed to dezenkerize tissues.

A
  • Lugol’s iodine
  • Sodium thiosulfate
74
Q

What is the fixation time for Heidenhain’s Susa?

A

3-12 hours

75
Q

What are the components of B-5 fixative?

A
  • Distilled water
  • Mercuric chloride
  • Anhydrous sodium acetate
76
Q

What is a significant disadvantage of using picric acid as a fixative?

A

Causes RBC hemolysis

77
Q

What is a key feature of Bouin’s solution?

A

Recommended for fixation of embryos and pituitary biopsies

78
Q

What is the primary disadvantage of using ethyl alcohol for fixation?

A

Destroys cytoplasmic structures

79
Q

What does the term ‘microanatomic fixative’ refer to in histology?

A

A fixative that preserves tissue structure for microscopic examination

80
Q

What is the main characteristic of compound fixatives?

A

Always correlated with other fixatives like Carnoy’s, Bouin’s, and Heidenhain’s Susa.

81
Q

At what temperature does the compound fixative solidify?

A

17 °C

82
Q

What are the advantages of using formalin as a fixative?

A
  • Fixes and precipitates nucleoproteins
  • Precipitates chromosomes and chromatin materials
  • Essential constituent of most compound nuclear fixatives
  • Causes tissues containing collagen to swell
83
Q

What are the disadvantages of using formalin for cytoplasmic fixation?

A

Destroys mitochondria and Golgi elements of the cells.

84
Q

What is the function of alcoholic fixatives?

A

Denatures and precipitates proteins by destroying hydrogen and other bonds.

85
Q

What concentration range is required for alcoholic fixatives?

A

70-100%

86
Q

What are the pros of using alcoholic fixatives?

A
  • Ideal for small tissue fragments
  • Can be used as both a fixative and dehydrating agent
  • Excellent for glycogen preservation
  • Preserves nuclear stains
87
Q

What are the cons of using low concentrations of alcoholic fixatives?

A
  • May cause RBC hemolysis
  • Inadequately preserves leukocytes
  • Dissolves fats and lipids
  • Causes polarization of glycogen granules
88
Q

What is the fixation time for methyl alcohol 100%?

A

Prolonged fixation for more than 48 hours may overharden tissue.

89
Q

What is Carnoy’s fluid primarily used for?

A

Fixing chromosomes, lymph glands, and urgent biopsies.

90
Q

What are the components of Carnoy’s fluid?

A
  • Absolute alcohol
  • Chloroform
  • Glacial Acetic Acid
91
Q

What is the fixation time for Carnoy’s fluid?

A

1-3 hours

92
Q

What is the main advantage of using osmium tetroxide?

A

Fixes conjugated fats and lipids permanently.

93
Q

What are the disadvantages of osmium tetroxide?

A
  • Very expensive
  • Poor penetrating agent
  • Readily reduced by contact with organic matter
  • Prolonged exposure may cause blindness
94
Q

What is the purpose of secondary fixation?

A

To facilitate and improve the demonstration of particular substances.

95
Q

What factors slow down the fixation of tissues?

A
  • Size and thickness of tissue specimen
  • Presence of mucus
  • Presence of fat
  • Presence of blood
  • Cold temperature
96
Q

What factors enhance the fixation of tissues?

A
  • Size and thickness of tissues
  • Agitation
  • Moderate heat (37-56 °C)
97
Q

What is the process of dehydration in tissue preparation?

A

Removing intracellular and extracellular water from the tissue after fixation.

98
Q

What are commonly used dehydrating agents?

A
  • Alcohol
  • Ethanol/Ethyl Alcohol
  • Isopropanol/Isopropyl Alcohol
  • Methanol/Methyl Alcohol
  • Butanol/Butyl Alcohol
  • Dioxane/Diethylene Dioxide
99
Q

What is the ideal volume of dehydrating agent needed?

A

Not less than 10x the volume of tissue to be dehydrated.

100
Q

What is the purpose of clearing in tissue preparation?

A

To replace the dehydrating agent with a fluid that will dissolve the wax.

101
Q

What is a common clearing agent?

A

Xylene

102
Q

What is the characteristic of xylene as a clearing agent?

A

It is the most rapid clearing agent.

103
Q

What is the definition of decalcification?

A

Technique for removing minerals from bone or other calcified tissue.

104
Q

What can overcalcification lead to?

A

Hard overstaining of tissues.

105
Q

What is the purpose of decalcification in tissue processing?

A

To remove calcium from tissues for examination, particularly in bones and associated pathologies

Decalcification is essential for diagnosing tumors, infections, and analyzing degenerative processes.

106
Q

What are some common tissues that may undergo decalcification?

A

Tissues undergoing degenerative processes, including:
* Bone
* Bone marrow
* Blood vessel walls
* Kidney
* Lung
* Areas with metastatic calcification

These tissues may exhibit calcium deposits that require removal for proper analysis.

107
Q

What is a good decalcifying agent characterized by?

A

Must be capable of:
* Completely removing calcium salts
* Not significantly damaging cells and tissue components
* Preserving staining capacity of the nucleus

Effective decalcifying agents maintain tissue integrity while allowing for microscopic analysis.

108
Q

List four types of agents used for calcium removal during decalcification.

A
  • Acids
  • Chelating agents
  • Ion exchange resins
  • Electrical ionization (electrophoresis)

Each type has different applications and effects on tissue morphology.

109
Q

What factors influence the rate of decalcification?

A
  • Concentration of active agent
  • Temperature
  • Agitation
  • Fluid access

Each factor can impact the efficiency and effectiveness of the decalcification process.

110
Q

True or False: Higher concentrations of decalcifying agents always result in less tissue damage.

A

False

Higher concentrations can increase tissue shrinkage and morphological damage.

111
Q

Name three commonly used acid decalcifying agents.

A
  • Nitric acid
  • Hydrochloric acid
  • Formic acid

These agents vary in effectiveness and impact on tissue morphology.

112
Q

What is the main disadvantage of using nitric acid as a decalcifying agent?

A

Inhibits nuclear stains and can destroy tissues, especially in concentrated solutions

This limits its application for certain diagnostic purposes.

113
Q

Fill in the blank: The most rapid decalcifying agent recommended for urgent works is _______.

A

Phloroglucin nitric acid

It allows for quicker processing times but may lead to poor nuclear staining.

114
Q

What is the role of EDTA in decalcification?

A

It acts as a chelating agent that combines with calcium ions to facilitate their removal

While effective, it is slow-acting and requires a large volume.

115
Q

What is the purpose of the impregnation process in histology?

A

To fill natural cavities and spaces in tissues with a medium that allows cutting of thin sections

This process is crucial for maintaining tissue structure during sectioning.

116
Q

What is a common method for impregnation in histology?

A
  • Paraffin wax impregnation
  • Celloidin wax/Collodion impregnation
  • Gelatin impregnation

Each method has specific applications based on the nature of the tissue being processed.

117
Q

What is the significance of maintaining the temperature of paraffin during embedding?

A

It must be maintained 2-5°C above the melting point to prevent tissue shrinkage and hardening

Proper temperature control is essential for optimal embedding.

118
Q

What is the recommended reagent to tissue ratio for fixation?

A

20:1

This ensures adequate fixation of the tissue specimen.

119
Q

What is the process of embedding in histology?

A

The process of placing impregnated tissue into a mold containing an embedding medium to solidify

Proper embedding is crucial for maintaining tissue orientation and integrity during sectioning.

120
Q

What is a potential consequence of improper labeling in a laboratory setting?

A

Mislabeling can lead to inaccurate results and jeopardize patient diagnosis

Accurate labeling is essential for maintaining the integrity of laboratory processes.

121
Q

What is the purpose of using cold water in tissue processing?

A

To solidify tissues, giving them a firmer consistency and better support for section cutting.

This facilitates the cutting of sections.

122
Q

What is a cold plate used for in tissue processing?

A

To easily solidify the embedding mold.

It aids in the hardening process of the tissue embedding.

123
Q

What is the purpose of a forceps warmer?

A

To properly arrange or have a proper orientation of tissue samples during manipulation.

124
Q

What is a paper boat used for in tissue embedding?

A

For embedding celloidin and paraffin blocks.

125
Q

What is the celloidin/nitrocellulose method recommended for?

A

Embedding hard tissues such as bones and teeth and large sections of whole organs like eyes.

126
Q

What is Low Viscosity Nitrocellulose (LV.N)?

A

A form of celloidin soluble in equal concentrations of ether and alcohol.

127
Q

What are the types of blocking-out molds mentioned?

A
  1. Leukhart’s embedding mold
  2. Compound embedding mold
128
Q

What does the double embedding method involve?

A

Tissues are infiltrated with celloidin and subsequently embedded in paraffin mass.

129
Q

What is the plastic or resin method especially good for?

A

Light microscopic studies, particularly in hard tissues and high-resolution light microscopy.

130
Q

What are some types of plastics used in embedding?

A

Epoxy plastics, Polyester plastics, Acrylic plastics.

131
Q

What is the purpose of microtomy?

A

To trim and cut processed tissues into uniformly thin slices for microscopic studies.

132
Q

What is trimming in the context of microtomy?

A

Cutting off excess wax from the block to expose the tissue surface.

133
Q

What is sectioning or section cutting?

A

Cutting tissues into uniformly thin slices with the aid of a machine.

134
Q

What is a microtome?

A

A machine/instrument designed for accurately cutting thin slices of tissues.

135
Q

What is the most common solution used during frozen sectioning?

A

Liquid nitrogen.

136
Q

What does honing involve?

A

The removal of gross nicks and irregularities on the knife edge.

137
Q

What is the direction used during honing?

A

Heel to toe (edge first).

138
Q

What is a hone?

A

A natural sharpening stone or hard grinding surface for sharpening knives.

139
Q

What are the types of microtomes mentioned?

A
  1. Sliding microtome
  2. Rotary microtome
  3. Rocking microtome
  4. Freezing microtome
  5. Ultrathin microtome
140
Q

What is a sliding microtome used for?

A

Cutting celloidin embedded sections.

141
Q

What is the most popular type of microtome?

A

Rotary microtome.

142
Q

What is the purpose of a freezing microtome?

A

For cutting unembedded frozen sections.

143
Q

What are the types of hones?

A
  1. Carborundum hone
  2. Arkansas stone
  3. Yellow Belgian/Belgium hone
144
Q

What is stropping?

A

The process of removing debris from the knife edge and polishing it.

145
Q

What is the direction used during stropping?

A

Toe to heel (edge last).

146
Q

What is the purpose of a floating-out bath?

A

To flatten sections and visualize folds and creases in sections.

147
Q

What is a cryostat used for?

A

For rapid preparation of urgent tissue biopsies and fluorescent antibody staining techniques.

148
Q

What are the two methods widely used for staining cryostat sections?

A
  1. H&E staining technique
  2. Polychrome methylene blue.
149
Q

What are frozen sections used for?

A

Rapid diagnosis of tissue and demonstration of lipids and nervous tissue elements.

150
Q

What are common methods of freezing for frozen sections?

A
  1. Liquid nitrogen
  2. Isopentane cooled by liquid nitrogen
  3. Carbon dioxide gas
  4. Aerosol sprays.
151
Q

What is freeze drying?

A

Preserving tissues by rapid freezing and removing water without chemical fixatives.

152
Q

What is the advantage of freeze drying?

A

Produces minimum shrinkage and allows tissues to be processed in a fresh state.

153
Q

What is freeze substitution?

A

A process similar to freeze-drying but involves fixing tissues before dehydration.

154
Q

What is Rossman’s fluid composed of?

A

Saturated Picric acid and Formaldehyde.

HISTOPATHOLOGY (2 decks)

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