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Histopathology and Cytopathology > E&M, Cytologic Techniques, Tissue Processing > Flashcards

E&M, Cytologic Techniques, Tissue Processing Flashcards

1
Q

(from the Greek mikros, meaning “small”, and temnein, meaning “to cut”) is a tool used to cut extremely thin slices of material, known as sections.

A

Microtome

The rotary microtome is the most used microtome in the Histopathology laboratory.

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2
Q

_____ is an instrument that is used to analyze and process samples by fixing, staining, dehydrating, or decalcifying them. It removes excess water from the tissue for it to be able to coat in melted paraffin wax.

A

Automatic tissue processor

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3
Q

_____ is designed to ensure that reagent solution spread evenly over the slide, and remain in place during the ensuing incubation period.

A

Automatic slide staining machine

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4
Q

It is designed for high-precision temperature control of embedding mediums like paraffin wax.

A

Paraffin oven

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5
Q

_____ is an essential step in the histology process, which allows the paraffin ribbon to smooth out and stick to the glass slide when placed.

A

Water bath

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6
Q

_____ is designed to assist with the handling of paraffin wax samples in histology and pathology laboratories. It is essentially a hot distilled water floating out bath that allows for the meticulous manipulation and location of sections onto glass slides.

A

Paraffin embedding wax bath

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7
Q

_____ is a rotor-driven instrument designed to separate liquid samples at high rates of speed.

A

Centrifuge

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8
Q

_____ is an appliance for removing excess paraffin from tissue cassettes

A

Paraffin block trimmer

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9
Q

_____ is designed to rapidly dry slides by blowing warm air over the surface of the slides.

A

Slide dryer

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10
Q

_____ is used to visualize very minute objects such as cells, microorganisms, giving a contrasting image, that is magnified.

A

Microscope

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11
Q

95% alcohol

-Pap smear staining-

A

Used in fixation (15-30 min)

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12
Q

80% alcohol

-Pap smear staining-

A

Used in hydration (2 min)

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13
Q

60% alcohol

-Pap smear staining-

A

Used in hydration (2 min)

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14
Q

1st Coplin jar of distilled water

-Pap smear staining-

A

Used in hydration (5 dips)

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15
Q

2nd Coplin jar of distilled water

-Pap smear staining-

A

Used in hydration (5 dips)

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16
Q

Hematoxylin stain

-Pap smear staining-

A

1st nuclear stain (3 min)

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17
Q

Running water

-Pap smear staining-

A

Gentle washing (30 sec)

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18
Q

Scott’s tap water

-Pap smear staining-

A

Blueing agent, pH=8.02 (3 min)

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19
Q

Distilled water

-Pap smear staining-

A

Dehydration (3 min)

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20
Q

60% alcohol

-Pap smear staining-

A

Dehydration (2 min)

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21
Q

80% alcohol

-Pap smear staining-

A

Dehydration (2 min)

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22
Q

1st Coplin jar of 95% alcohol

-Pap smear staining-

A

Dehydration (2 min)

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23
Q

Orange G stain

-Pap smear staining-

A

1st counterstain (3 min)

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24
Q

1st Coplin jar of 95% alcohol

-Pap smear staining-

A

(2 min)

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25
Q

2nd Coplin jar of 95% alcohol

-Pap smear staining-

A

(2 min)

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26
Q

Eosin Azure stain

-Pap smear staining-

A

2nd counterstain (3 min)

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27
Q

1st Coplin jar of 95% alcohol

-Pap smear staining-

A

(2 min)

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28
Q

2nd Coplin jar of 95% alcohol

-Pap smear staining-

A

(2 min)

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29
Q

1st Coplin jar of absolute alcohol

-Pap smear staining-

A

(2 min)

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30
Q

2nd Coplin jar of absolute alcohol

-Pap smear staining-

A

(2 min)

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31
Q

Step in tissue processing prior to fixation

A

Obtaining a fresh specimen

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32
Q

_____ is used to preserve cells and tissue components in a “life-like state” or as little alteration as possible to the living tissue.

A

Fixation

Uses formalin/formaldehyde as a fixative or phosphate-buffered solution

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33
Q

_____ is the process of removing calcium from tissues

A

Decalcification

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34
Q

_____ is simply the removal of water from aqueous-fixed tissue. This step is necessary to prepare the tissue for embedding in non-aqueous media like paraffin.

A

Dehydration

Uses increasing concentrations of ethanol solutions to avoid excessive distortion of tissue

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35
Q

The purpose of _____ is to remove dehydrating agents from tissues and to prepare the tissues for impregnation with the embedding agent. It also removes a substantial amount of fat from the tissue.

A

Clearing

Uses xylene immersions (1 to 3 kinds) for 5 min each

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36
Q

The role of this agent is to remove the clearing agent from the tissue and to completely permeate the tissue with paraffin wax. This step is called?

A

Infiltration/Impregnation

Uses melted paraffin wax (usually at 60 C, then allowed to cool at 20 C to solidify). These waxes are a mixture of purified paraffin wax and resins such as styrene or polyethylene.

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37
Q

_____ is the process in which the tissues or the specimens are enclosed in a mass of the embedding medium using a mold (cassette).

A

Embedding

Epoxy resins are the most commonly used embedding media for semi-thin and ultrathin sections.

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38
Q

_____ is cutting a fixed tissue or organ to create a flat surface with the correct orientation.

A

Trimming

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39
Q

_____ is the technique of making very thin slices of tissue specimens for microscopic examination.

A

Section cutting/Microtomy

Specimens for H&E routine staining are cut 3-5 micrometers in thickness.

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40
Q

Parafinized ribbons are removed from the microtome using a wooden tongue depressor. Uses albumin and poly-l-lysine additives as a bonding agent.

A

Mounting

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41
Q

After drying at 60 C, the slide is passed through another series of chemical reagents.

A

Staining

Uses xylene to remove paraffin, then uses absolute alcohol to remove xylene.

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42
Q

It affects infiltration and subsequent sectioning of tissues.

A

Tissue density and thickness

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43
Q

It increases the flow of fresh fluids around the tissue.

A

Agitation

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44
Q

It can speed up fluid penetration and tissue processing protocols

A

Temperature (37 C-45 C)

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45
Q

Reduced pressure can increase the infiltration rate and decrease the time needed to complete the steps in tissue processing protocols.

A

Vacuum and Pressure

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46
Q

_____ is a diagnostic technique used to examine cells from body fluids and solid tissues to determine the nature of the disease.

A

Cytology

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47
Q

_____ is the microscopic study of normal tissues of the body. While _____ is the microscopic study of the tissues affected by the disease.

A

Histology

Histopathology

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48
Q

Cells shed from bodily surfaces, such as the inside of the mouth, are collected and analyzed.

A 
Exfoliative Cytology (Ex: urine, sputum, CSF, effusions, 
etc.)
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49
Q

Cells are obtained directly from the surface of the target of interest or from dislodged cells from the body’s surfaces or mucosal linings.

A

Abrasive Cytology (Ex: Scrapings from the cervix (pap smear), vagina, oral cavity (buccal mucosal smear), and skin lesions, bronchial wash and swabs)

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50
Q

____ is a technique used for obtaining material from organs that do not shed cells on their own. It is also called Fine Needle Aspiration Cytology (FNAC) because it uses a fine needle to obtain samples.

A

Aspiration Cytology (Ex: subcutaneous soft tissue, tumors, thyroid, lymph nodes, salivary glands, and breast)

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51
Q

It is a polychromatic stain that contains multiple dyes to differentially stain various components of the cell and it is considered the most common stain used in cytopathology.

A

Papanicolaou stain (PAP smear stain)

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52
Q

This approach is used to distinguish cells in the smear preparation of various gynecological specimens (pap smears) and materials comprising exfoliative cytology smears.

A

Pap smear staining

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53
Q

This technique is useful in lesions that are easily palpable. For non-palpable, it requires the use of ultrasound or CT guidance.

A

Aspiration Cytology

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54
Q

_____ are used to differentiate cells for microscopic examination in pathological specimens. Used in FNACs, and fluids

A

Romanowsky stains

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55
Q

Type of Romanowsky stains

A

Leishman’s stain - staining blood smears
Wright’s stain - staining peripheral blood smears, urine samples, and bone marrow aspirates
Giemsa’s stain/May Grunwald Giemsa - gold standard staining technique that is used for both thin and thick smears
Jenner’s stain
Field’s stain

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56
Q

_____ stain is also used in FNACs and fluids. Mainly used as histopathology stain.

A

H&E (Hematoxylin and Eosin)

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57
Q

This is the most common and easiest preparation of the cellular material.

A

Direct smear

58
Q

Another method of preparation of the cellular materials are:

A

Cytospin preparation
Membrane filter preparation
Liquid-based preparation
Cell blocks

59
Q

Air-dried fixation is done to smears that use what stain?

A

MGG/May Grunwald Giemsa/Giemsa

60
Q

Air-dried fixation is later wet-fixed in ______ for 15-20 mins during the staining process

A

Methyl-alcohol

61
Q

______ done using cytosprays is employed in many labs as an easier alternative to wet fixation

A

Spray fixation

62
Q

The ideal angle for spray fixation is ____

A

45 degrees

63
Q

Ideal distance for spray fixation is _____

A

15-25 cm

64
Q

The fixative agent that is held too close, causes ____

A

dislodgement of the cells from the smear

65
Q

The fixative agent that is held too far, causes ____

A

insufficient amount of fixative

66
Q

The fixative agent that is forcefully sprayed, causes _____

A

creates cellular artifacts by the fracturing of cytoplasm

67
Q

Father of Cytopathology

A

George Papanicolaou

68
Q

_____ stains basic component of cells

A

Acid dye

69
Q

_____ stains acid component of cells

A

Basic dye

70
Q

Nuclei are stained by _____

A

hematoxylin solution

71
Q

This stain is used as a cytoplasmic stain

A

Orange G stain

72
Q

A polychromatic solution contains a mixture of _____ that is responsible for the demonstration of differentiation of squamous cells

A

Eosin Y
Light Green SF
Bismarck Brown Y

73
Q

It stains the cell nuclei blue.

A 
Harris Hematoxylin (most common)
Gill's Hematoxylin and Hematoxylin S (other stains)
74
Q

It is the first acidic counterstain (cytoplasmic stain) which stains mature and keratinized cells

A

Orange Green 6 (OG-6)

75
Q

It is the second counterstain which is a polychrome mixture of Eosin Y, Light Green SF, Bismarck Brown. It gives pink color to the cytoplasm of mature squamous cells, nucleoli, cilia, and red blood cells.

A

Eosin Azure 50 (EA-50)

76
Q

It stains blue to cytoplasms of cells like parabasal squamous cells, intermediate squamous cells, and columnar cells.

A

Light Green SF

77
Q

It stains nothing instead it stabilizes the staining.

A

Bismarck Brown Y

78
Q

Three types of PAP stain:

A

Progressive
Regressive
Rapid

79
Q

The nucleus is stained with hematoxylin to an intensity desired

A

Progressive method of PAP stain

80
Q

The nucleus is deliberately over-stained with a non-acidified hematoxylin.

A

Regressive method of PAP stain

81
Q

Color of stain: Nuclei

A

Blue

82
Q

Color of stain: Acidophilic cells

A

Red

83
Q

Color of stain: Basophilic cells

A

Blue-green

84
Q

Color of stain: Erythrocytes

A

Orange-red

85
Q

Color of stain: Keratin

A

Orange-red

86
Q

Color of stain: Superficial cells

A

Pink

87
Q

Color of stain: Intermediate and Parabasal cells

A

Blue-green

88
Q

Color of stain: Eosinophils

A

Orange-red

89
Q

List of Precautions:

A

Immediate fixation of smear is essential
Solutions and other stains are filtered daily after use to keep them free of sediment
Alcohols are hygroscopic
Stains are discarded and replaced as the quality of the stain deteriorates
Place the coverslip on the microslide slowly without trapping air bubbles
Stains keep longer if they are stored in dark-colored stoppered bottles

90
Q

When alcohol is left open, the concentration _____

A

decreases

91
Q

Blueing solution and HCl should be replaced _____

A

once daily

92
Q

Water rinses should be changed _____

A

after use

93
Q

Alcohol used for the process of dehydration prior to the cytoplasmic stains should be replaced _____

A

weekly

94
Q

Accurate patient identification, orientation of samples and adequate fixation rest are responsibilities of _____.

A

OT staff

95
Q

Appropriate grossing, sampling are responsibilities of ____.

A

Junior pathology staff

96
Q

Adequate processing, appropriate embedding techniques, microtomy, staining, and avoiding unacceptable artifacts are responsibilities of _____.

A

Histopathology technicians

97
Q

Inspection of control, immuno-histochemical methods, and reporting of the slides are all responsibilities of _____.

A

Expert pathologist

98
Q

Tissues are collected and fixed in _____.

A

10% formalin

99
Q

Stain for testicular biopsy.

A

Bouin’s solution

100
Q

Test requisition form should contain:

A 
patient demographics
detail of patient's history
anatomic site of biopsy
number of containers sent
specimen transportation instruction etc.
101
Q

Specimens are transported at what temperature?

A

room temperature

102
Q

Specimen for transport must be labeled with what?

A

10% formalin hazard label

103
Q

Specimen for transport must be placed in what?

A

well-sealed leak-proof container

104
Q

Specimen identification is maintained across steps like:

A

Specimen labeling
Grossing
Block labeling
Slide labeling etc.

105
Q

Bony or cartilaginous tissue are placed in decalcifying solution for how many days?

A

1-7 days

106
Q

The tissue is washed _____ times in distilled water

A

3-4

107
Q

Grossing station must be cleaned with _____ daily after grossing.

A

1% sodium hypochlorite

108
Q

Formalin is volatile and toxic and causes irritation to what parts of the body?

A

eyes, mucus membranes, skin

109
Q

Define OSHA:

A

Occupational Safety and Health Administration

110
Q

average formalin exposure?

A
  1. 75 ppm / 8hr

2. 0 ppm / 15 minutes (short-term)

111
Q

Alcohol concentrations in Dehydration:

A 
50% alcohol - 90 min
70% alcohol - 90 min
80% alcohol - 90 min
95% alcohol - 90 min
100% alcohol - 90 min
100% alcohol - 12 hr
112
Q

Clearing agent used is ____

A

Xylene

113
Q

It is the simplest, least invasive test and uses the smallest needle to simply remove cells from the area of abnormality.

A

Fine Needle Aspiration/Fine Needle Aspiration Cytology

114
Q

It removes not only cells but also a small portion of the surrounding tissue. This provides additional information to assist in the examination of the lesion.

A

Core needle biopsy

115
Q

It takes out even more surrounding tissues. It takes some of the abnormality but not all. This process involves slicing into the lesion but removing only a portion of it.

A

Incisional biopsy

116
Q

It generally removes the entire area in question.

A

Excisional biopsy

117
Q

It is considered the primary technique for obtaining diagnostic full-thickness skin specimens. This technique involves the use of a circular blade that is rooted down through the epidermis and the dermis, and into the subcutaneous fat, yielding a 3-4 mm cylindrical core of the tissue sample.

A

Punch biopsy

118
Q

It is where all small fragments of tissue are shaved from a surface (usually a skin)

A

Shave biopsy

119
Q

It is where tissue is scooped or spooned to remove tissue or growths from the body cavity such as the endometrium or cervical canal.

A

Curettings

120
Q

It is a process whereby a selected tissue specimen is immersed in isotonic salt solution such as normal saline solution or Ringer’s solution in a petri dish or watch glass, carefully dissected with a needle, and separated by direct or zigzag spread using an applicator stick

A

Teasing or dissociation

121
Q

It is a process whereby small pieces of tissue are placed in a microscopic slide and forcibly compressed with another slide or with a cover glass.

A

Squash preparation/Crushing

122
Q

With an applicator stick or a platinum loop, the material is rapidly and gently applied in a direct or zigzag line throughout the slide, attempting to obtain a uniform distribution of secretion.

A

Streaking (Smear preparation)

123
Q

A selected portion of the material is transferred into the clean slide and gently spread into moderately thick film by teasing the mucus strands with an applicator stick. It is specially recommended for smear preparation of fresh sputum and bronchial aspirate, and also for thick mucoid secretions.

A

Spreading (Smear preparation)

124
Q

This is done by placing a drop of secretion or sediment upon one slide and facing it unto another clean slide.

A

Pull-apart (Smear preparation)

125
Q

This is a special method of smear preparation whereby the surface of a freshly cut piece of tissue is brought into contact and pressed on the surface of a clean glass slide.

A

Touch Preparation/Impression smear

126
Q

Immediate diagnosis is accomplished through the use of ______

A

Frozen section

127
Q

Fresh tissue is frozen on a microtome with CO2 or on a ______, which is a cold chamber kept at an atmospheric temperature of -10 to -20 C.

A

cryostat

128
Q

It is generally used in histochemistry. It is the most rapid of the commonly available freezing agents.

A

liquid nitrogen

129
Q

The main disadvantage of using liquid nitrogen as a freezing agent is:

A

It produces ice crystals or freezes artifacts on the tissue

130
Q

A freezing agent that is liquid at room temperature

A

isopentane

131
Q

Other freezing agents are:

A

CO2 gas and aerosol sprays (cryokwik)

132
Q

It is a refrigerated apparatus used for fresh tissue microtome which maintains the temperature between -5 to -30 C (the average is -20 C)

A

Cryostat/Cold microtome

133
Q

It is capable of freezing fresh tissues within 2-3 minutes

A

Thermostat

134
Q

It is a special way of preserving tissues by rapid quenching of fresh tissue at -160 C and subsequently removing ice molecules without using any fixative.

Time-consuming and expensive

A

Freeze drying/Rapid freezing

135
Q

vacuum at -40 C

A

Sublimation

136
Q

It is a process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient temperature dehydration.

More economical and less time consuming

A

Freeze substitution

137
Q

Fixative for frozen tissue:

A

Rossman’s Formula

1% acetone

138
Q

Reagent for dehydration of tissue:

A

Absolute alcohol

139
Q

Reagent change:

A

Absolute alcohol - alternate day
Xylene - every 3rd day
Wax - weekly
Formalin - every day

140
Q

A special stone used to sharpen the knife in a microtome is called _____. The process of sharpening is called _____

A

Hone

Honing

141
Q

It is the process of polishing an already fairly sharp edge of the knife that may be flexible or rigid. The material is called _____.

A

Stropping

Strop

142
Q

Labeling is an important step and must be done using _____

A

diamond pencil

Histopathology and Cytopathology (2 decks)

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