PRELIM LAB: THE HISTOPATHOLOGY LABORATORY Flashcards

- HISTPATHOLOGY LABORATORY, EXAMINATION OF TISSUES IN HISTOPATHOLOGY LABORATORY, FIXED TISSUE EXAMINATION

1
Q

Study of tissues affected by disease

fdf

A

HISTOPATHOLOGY

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2
Q

Useful in making a diagnosis and in determining the severity and progress of a condition

A

HISTOPATHOLOGY

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3
Q

Duration of specimen storage:

A

at least 1-2 weeks

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4
Q

obsolete, not often used
nowadays

A

Fresh tissue examination

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4
Q

should be stored in the
lab forever

A

Autopsy specimen

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5
Q

head of the laboratory

A

Pathologist

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6
Q

supports the pathologist

A

Associate Pathologist

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6
Q
  • The medical technologist
  • Provides slides that are properly labeled,
    processed, stained, mounted, and
    sequenced
A

Histotechnologist/Histotechnician

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7
Q
  • Ensures that formalin and other agents are
    fresh and in good working quality
  • Maintains equipment in high-quality condition
  • Performs preventing maintenance, as well as
    troubleshooting procedures
A

Histotechnologist/Histotechnician

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8
Q
  • Sections large and hollow organs to allow fixation
  • Examines the tissue sections, cytologic slides
    under the microscope
  • Monitors staff performance
  • Pinpoint problematic situations and find
    solutions
A

Pathologist and Assistant Pathologist

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9
Q

Set of procedures or technical activities on fulfilling
quality

A

QUALITY CONTROL

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10
Q
  • Ensuring that everything is right (test, time,
    specimen, patient, diagnosis, and price)
  • Includes availability of reagents, supplies, preventive maintenance and monitoring of equipment and evaluation of the quality of services
A

QUALITY ASSURANCE

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11
Q

TYPES OF HISTOPATHOLOGIC PROCESS

A
  1. Tissue Processing
  2. Frozen Biopsy
  3. Special Staining
  4. Immunohistochemistry
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12
Q

HISTOPATHOLOGIC TECHNIQUES

A

Includes all activities done in the laboratory in order to produce a suitable specimen side for viewing by the pathologist

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13
Q
  • Proper specimen collection and processing of results and documentation
  • High quality of reagents and equipment
  • Continuous professional education of staff
A

QUALITY MANAGEMENT SYSTEMS

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14
Q
  • Set of coordinated activities to regulate a lab in order to continually improve its performance
  • Skilled personnel
  • Considers pre-analytic, analytic, and post-analytic phase
A

QUALITY MANAGEMENT SYSTEMS

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15
Q

from receiving of specimen to
encoding of patient information

A

Pre-analytic

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16
Q

Skilled histotechnologist/histotechnician
which are responsible for:

A
  • Proper specimen collection
  • Proper processing of specimen
  • Efficient processing of results
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16
Q

tissue processing phase

A

Analytic

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17
Q

To ensure an effective QMS, the histopathology laboratory should have the following:

A

Skilled histotechnologist/histotechnician

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17
Q

reading of slides and final
diagnosis

A

Post-analytic

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18
Q
A
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19
Q

Numerically, alphabetically, or chronologically arranged

A

DOCUMENTS

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20
Q

Request Form

A
  1. Name, Age, Sex, Date of Birth
  2. Hospital or Lab Accession #
  3. Specimen Type/Source; Clinical Impressions
  4. Pertinent History, Operative Findings
  5. Test Requested, Procedure performed
  6. Date & Time of Request, Collection, and
    Transport
  7. Requesting Physician
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21
Q

Request Form info + Diagnosis and
Gross/Microscopic findings + Name of
pathologist

A

Patient Report

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22
Q

TYPES OF RESULTS

A
  1. Surgical Pathology
  2. Cytopathology
  3. Autopsy Report
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23
Q

TURNAROUND TIME (TAT) OF RESULTS

Surgical Pathology and Cytology

A

2 days

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24
Q

TURNAROUND TIME (TAT) OF RESULTS

Frozen Sections

A

5-15 minutes

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25
Q

TURNAROUND TIME (TAT) OF RESULTS

Autopsy Report

A

7 days

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26
Q

preliminary diagnosis

A

Telephone Report

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27
Q

status of results 48-74 hours from
receiving

A

Prelim Report

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28
Q

conveys results after test is completed

A

Final Report

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29
Q
A
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30
Q

documents occurrence of problems

A

Incident Report

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31
Q

Tissues for examination are usually obtained through biopsy or autopsy. They range from whole organs or very large specimens to tiny fragments of tissue

A

EXAMINATION OF TISSUES IN HISTOPATHOLOGY LABORATORY

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32
Q

● AKA Necropsy; Thanatopsy
● Post-mortem examination of tissues

A

AUTOPSY

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33
Q

AUTOPSY PURPOSES:

A

● Determine cause of death and extent of injury
● Uncovering existence of an undetected disease

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34
Q

TYPES OF AUTOPSY ACCORDING TO:

A
  • PURPOSE
  • COMPLETENESS
  • MANNER OF INCISION
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35
Q

TYPES OF AUTOPSY ACCORDING TO: PURPOSE

performed on a patient
who dies in a hospital during course of treatment

A

Medical/Hospital

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36
Q

TYPES OF AUTOPSY ACCORDING TO:

● Partial
● Complete

A

COMPLETENESS

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37
Q

TYPES OF AUTOPSY ACCORDING TO:

● Y-shaped
● Straight Cut (I-shaped)

A

MANNER OF INCISION

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38
Q

TYPES OF AUTOPSY ACCORDING TO: PURPOSE

generates evidentiary document that forms a basis for opinions rendered in a criminal trial, civil suit, and the like

A

Medico-legal

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39
Q

DISSECTION/EVISCERATION TECHNIQUES

A
  • VIRCHOW
  • ROKITANSKY
  • GHON
  • LETULLE
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40
Q

DISSECTION/EVISCERATION TECHNIQUES

● One by one removal of organs
● MOST WIDELY USED

A

VIRCHOW

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41
Q

DISSECTION/EVISCERATION TECHNIQUES

● “In situ” (in place) dissection, followed by en bloc removal

A

ROKITANSKY

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42
Q

DISSECTION/EVISCERATION TECHNIQUES

● “en bloc” removal
● Organs of same group/cavity/region are removed at the same time

A

GHON

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43
Q

DISSECTION/EVISCERATION TECHNIQUES

● “en masse” removal of organs
● All organs are removed at the same time, then dissected by blocks

A

LETULLE

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44
Q

PREREQUISITES TO PERFORMING AUTOPSY

A
  1. Written or informed consent from the legal next-of-kin
    - Order of priority: spouse, adult child, either
    parent, adult sibling, grandparent, guardian
  2. Medical abstract or clinical data
  3. Autopsy Request (suspicious evidence of foul play)
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45
Q

PREREQUISITES TO PERFORMING AUTOPSY

spouse, adult child, either
parent, adult sibling, grandparent, guardian

A

Order of priority

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46
Q

PERSONNEL

A
  1. Coroner
  2. Prosector
  3. Diener
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47
Q

a public official who is empowered to order
an inquest into the manner or cause of death

A

Coroner

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48
Q

pathologist who performs the dissection

A

Prosector

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49
Q

comes from German word “leichendiener”
meaning “servant of the dead”; assists during
autopsy, and assumes many and varied
responsibilities in the autopsy laboratory

A

Diener

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50
Q

Organ block removed from the body cavity should be thoroughly washed of blood using ___________ to minimize the blood staining of organs. NEVER USE HOT WATER.

A

cool or cold water

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51
Q

Organ blocks are placed in a large enameled potmcontaining fixative, filled to about __________.

A

⅓ capacity

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52
Q

Tissues should not be pressed against each other ormthe bottom or walls of the container. TRUE OR FALSE?

A

TRUE

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53
Q

Lesions that are encountered during dissection should be _____________ before organ is fully incised.

A

obtained early and placed in fixative

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54
Q

● Ante-mortem examination of tissues (ante=before; mortem=death)
● Examination of tissue sample from the living

A

BIOPSY

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55
Q

TYPES OF BIOPSY

A
  1. FINE NEEDLE ASPIRATION (FNA)
  2. CORE NEEDLE
  3. INCISIONAL
  4. EXCISIONAL
  5. PUNCH
  6. SHAVE
  7. CURETTAGE
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56
Q

Simplest, LEAST INVASIVE
Uses very thin needle attached to syringe to take out small amount of fluid and tissue from area

A

FINE NEEDLE ASPIRATION (FNA)

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57
Q

Surgical; small part of a large lesion or tumor is taken

A

INCISIONAL

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57
Q

Uses slightly larger needle
Remove small column of tissue (1/16 inch in
diameter, ½ inch long)

A

CORE NEEDLE

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58
Q

Surgical; entire affected area is taken

A

EXCISIONAL

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59
Q

For skin; uses circular blade to obtain deeper skin sample that removes a short cylindrical core of tissue (“apple core”)

A

PUNCH

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60
Q

For skin; small fragments of out layers of skin are “shaved” or scraped

A

SHAVE

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61
Q

Tissues are removed from body cavity (or canals) using a curette (instrument with a tip shaped like a small scoop or hook)

A

CURETTAGE

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62
Q

METHODS OF EXAMINATION OF BIOPSY SPECIMENS

A
  1. FRESH
  2. PRESERVED
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63
Q

Allows examination of cells in their living state

A

FRESH TISSUE EXAMINATION

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64
Q

FRESH TISSUE EXAMINATION DISADVANTAGES

A

Subject to ischemia, therefore not
permanent, and liable to changes

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64
Q

FRESH TISSUE EXAMINATION ADVANTAGES

A

View protoplasmic activities (motion,
mitosis, phagocytosis, pinocytosis)

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65
Q

METHODS FOR FRESH TISSUE EXAMINATION

A
  1. Teasing
  2. Squash Preparation
  3. Smear
    a. Streaking
    b. Spreading
    c. Pull-apart
    d. Touch Prep
  4. Frozen Sections
66
Q

AKA Dissociation

A

TEASING

67
Q

Selected tissue is immersed in petri dish/watch glass containing isotonic solution, and then carefully dissected and separated using needle or applicated
stick

A

TEASING

68
Q

Tissue is then transferred to slide and examined under the microscope (phase contrast or bright field)

A

TEASING

69
Q

May be stained using supravital dyes

A

TEASING

70
Q

AKA Crushing

A

SQUASH PREPARATION

71
Q

Small pieces of tissue with diameter <1mm are
compressed between two slides

A

SQUASH PREPARATION

72
Q

May be stained using supravital dye

A

SQUASH PREPARATION

73
Q

Method depends on nature of material to be examined

A

SMEAR

74
Q

Useful for cytological examinations

A

SMEAR

75
Q

Cellular materials are spread over a slide using wire loop, applicator stick or slide

A

SMEAR

76
Q

Vital stains may also be applied

A

SMEAR

77
Q

May be made permanent by fixing them

A

SMEAR

78
Q

SMEAR TYPES:

A

a. Streaking
b. Spreading
c. Pull-apart
d. Touch Prep

79
Q

Rapid, but gentle zigzag application of the material throughout slide

A

Streaking

80
Q

Must have a relatively uniform distribution

A

Streaking

81
Q

Material is gently spread onto the slide, and
the mucus strands are teased apart using an
application stick

A

Spreading

82
Q

ADVANTAGE: Maintains intercellular relationships

A

Spreading

83
Q

For fresh sputum, bronchial aspirates, and
thick mucoid secretions

A

Spreading

84
Q

Slides are then pulled apart with a single,
uninterrupted movement in opposite
directions

A

Pull-apart

85
Q

A drop of the material is placed into a clean
slide and covered with another clean slide.
Material is allowed to disperse evenly

A

Pull-apart

86
Q

Surface of a freshly cut tissue is pressed to a clean slide

A

Touch Prep

86
Q

AKA Impression Smear

A

Touch Prep

87
Q

For Phase-Contrast Microscopy

A

Touch Prep

88
Q

ADVANTAGE: Maintains intracellular relationships

A

Touch Prep

89
Q

For rapid diagnosis of tissue

A

FROZEN SECTIONS

90
Q

Requested during intra-operative procedures to help surgeon in choosing his next plan of action

A

FROZEN SECTIONS

91
Q

FROZEN SECTIONS DISADVANTAGE:

A

relatively poor quality of the final
slide; expensive

91
Q

Fresh tissues are frozen using a

A

cryostat or freezing
microtome

91
Q

Recommended for demonstration of lipids and nervous tissue

A

FROZEN SECTIONS

92
Q

FROZEN SECTIONS ADVANTAGE:

A

rapid processing time with less equipment requirement and less need for ventilation

93
Q

Accomplished by fixing the tissues and carefully processing them to preserve their structures, then impregnating them with hardening substance to permit making thin slices suitable for staining and microscopic evaluation

A

FIXED TISSUE EXAMINATION

93
Q

● END GOAL: produce a tissue section of good quality that allows for adequate interpretation of microscopic cellular changes for diagnosis

A

FIXED TISSUE EXAMINATION

94
Q

```

Various steps required to take the tissue from fixation to the state where it is completely infiltrated with a suitable histological wax and can be embedded ready for section cutting on the microtome

A

TISSUE PROCESSING

95
Q

Preservation of tissue constituents in a life-like manner as possible

A

Fixation

96
Q

● Remove of calcium or lime salts from bones following fixation
● For ease of cutting

A

Decalcification (optional)

97
Q

Removal of water

A

Dehydration

98
Q

Removal of the dehydrating agent with the use of a clearing agent (a solvent miscible with both the dehydrant and impregnating material)

A

Clearing/Dealcoholization

99
Q

● Replacement of clearing material with impregnating material
● Gives firm consistency to tissue for ease of handling and sectioning

A

Infiltration/Impregnation

100
Q

Placing the infiltrated tissue into a mold filled with molten wax to form a solid tissue block

A

Embedding

101
Q

Removal of excess wax into fine tissue sections with the use of a microtome

A

Trimming

102
Q

Cutting of tissues into fine tissue sections with the
use of a microtome

A

Section-Cutting/Microtomy

103
Q

Application of dyes to sections for visualization

A

Staining

104
Q

Use of media to mount a coverslip for ease of
handling, storage, and protection of sectioned tissue

A

Mounting

105
Q

Before tissues are mounted on slides, they should undergo different processes. Each step in the process necessitates the use of various specific components.

A

INSTRUMENTATION

105
Q

Indicating of accession number and year for proper
identification

A

Labelling

106
Q

GROSS TABLE:

A

● Forceps
● Scalpel
● Chopping board
● Weighing balance
● Ruler
● Orientation markers
● Watch glass/petri dish
● Tissue cassettes
● Diamond pen, pencils

107
Q

AUTOMATED TISSUE PROCESSORS (FDCI)

A

● Paraffin oven
● Embedding center
● Refrigerator: 40C
○ Freezer: 20C

107
Q

Wax is heated at ____ in the embedding machine

A

60C

108
Q

Processes that an automated machine can perform

A

FDCI (Fixation, Dehydration, Clearing, Infiltration)

109
Q
  • For unstained and transparent samples
  • Only oblique rays hit the object
  • Samples are made brightly lit against dark
    background
A

Dark Field Microscope

109
Q
  • Phase shifts in light passing through transparent and colorless specimen are converted into brightness (contrast) changes in the specimen, making them visible
  • does not require staining
A

Phase Contrast Microscope

109
Q

for tissue and cell visualization

A

Microscope

110
Q

simplest and most popular

A

Bright Field Microscope

111
Q
  • Designed to examine birefringent properties of anisotropic specimens
  • Birefringence: splitting of one ray of light into two
  • Anisotropism: substances that exhibit different properties when measured in different directions
  • NOTE: Amyloid in Congo Red has Apple Green birefringence
A

Polarizing Microscope

112
Q
  • Uses ability of substance to exhibit fluorescence
  • Fluorescence: emission of low frequency light of substances when they are illuminated with high energy light
  • Only allows observation of the specific structures
    which have been labeled for fluorescence
  • NOTE: Acridine Orange stains ssDNA – green
    fluorescence, and ssDNA or RNA (cytoplasm) – red fluorescence;
A

Fluorescence Microscope

113
Q
  • Uses a beam of accelerated electrons as source of illumination
  • Has higher resolving power than light microscopes and can reveal the structure of smaller objects
A

Electron Microscope

114
Q

For producing tissue ribbons or sections

A

Microtome

115
Q

KINDS OF MICROTOME

A
  • Rotary
  • Cryostat
  • Sliding
  • Freezing
  • Rocking
  • Ultrathin
115
Q

Microtome Main Parts:

A

a. Block Holder
b. Knife Holder
c. Pawl and Feedwheel Mechanism

116
Q

MOST COMMONLY USED; for routine and
serial (continuous) sections; knife is stationary

A

Rotary

116
Q

Rotary Thickness:

A

3-5 micrometers

117
Q

Rotary Inventor:

A

Minot

117
Q

Rotary Microscope:

A

Light Microscope

118
Q

Rotary Embedding material:

A

Paraffin

118
Q

Sliding Types:

A

Base-Sledge and Standard Sliding

118
Q

consists of a microtome (usually rotary),
kept inside a cold chamber maintained at -5C to -30C (avg: -20C); capable of freezing fresh tissues, thus used for STAT diagnosis

A

Cryostat

119
Q

knife is moving

A

Sliding

120
Q

invented by Queckette (1848)

A

Freezing

120
Q

SIMPLEST

A

Rocking

121
Q

Sliding Inventor:

A

Addams (1783)

122
Q

60-100 nm thickness of tissue
- Mostly used for tissues embedded in epoxy
resin

A

Ultrathin

123
Q

best for antigen preservation

A

Microwave Oven

124
Q

used in epitope retrieval for immunohistochemistry

A

microwave oven

125
Q

eliminate the need for the laborious manual staining

A

automated stainers

126
Q

agitation and heating will increased fixation rate

A

microwave oven

127
Q
  • In situ hybridization and enzyme reactions
  • Thermal Requirements: 37C
A

Incubator Oven

128
Q

may operate through:
- dipping the slides into the stain; and/or
- applying the stain to the slides

A

automated stainers

129
Q

Eliminate the need for the laborious manual staining

A

automated stainers

129
Q

slide driers thermal requirement:

A

5-10C HIGHER than the
melting point of paraffin

129
Q

removing water collected during sectioning (water from floatation bath)

A

slide dryers

129
Q

causes uneven staining, artifacts
formation and tissue destruction

A

Overheating

130
Q

Fishing out of tissue section; keeps sections from
wrinkling

A

FLOTATION BATH

131
Q

Has “black” interior, for easy visualization of sections

A

FLOTATION BATH

132
Q

FLOTATION BATH Thermal requirement:

A

5-10C LOWER than the
melting point of paraffin

133
Q

System designed for paraffin embedding

A

EMBEDDING CENTERS

134
Q

EMBEDDING CENTERS Thermal requirement:

A

2-4C HIGHER than the
melting point of the paraffin

135
Q

MOST COMMON EMBEDDING CENTERS

A

PAPER BOAT

135
Q

EMBEDDING CENTERS Components:

A

○ Paraffin dispenser and reservoir
○ Orientation stage
○ Chilling plate
○ Warm storage (for embedding mold)

136
Q

● May be stationary or moving
● Equipped with alarm and warn technicians if high temperature
● Often use vacuum in heat to speed up procedures

A

AUTOMATED TISSUE PROCESSOR

137
Q

AUTOMATED TISSUE PROCESSOR MAIN TYPES:

A
  • Tissue-Transfer Machine (dip and dunk)
  • Fluid-Transfer Machine (enclosed)
138
Q

specimens are transferred from container to container

A

Tissue-Transfer Machine (dip and dunk)

139
Q

specimens are held in a single chamber, and then fluids are pumped in and out as required

A

Fluid-Transfer Machine (enclosed)

140
Q

For gross examination and dissection of submitted
specimens

A

GROSS TABLE

141
Q

GROSS TABLE It should have:

A

○ Sink
○ Table Top
○ Water Supply
○ Irrigation System
○ Fume Ventilation
○ Waste Disposal Unit

142
Q

Coplin jars, staining racks, staining dishes (all
three are for staining) and other glass and plastic materials (for storage and preparation of solutions)

A

Wares

143
Q

measurement of solution and buffers

A

pH Meters

144
Q

for gross examination

A

Grossing Tools

144
Q

Freezer Thermal requirement:

A

-20C

145
Q

Refrigerators Thermal requirement:

A

4C

146
Q

– liver tissues and renal
tissues

Tissue cassette

A

YELLOW

147
Q

routine tissues
(fallopian tube, cervix,
gallbladder, appendix)

Tissue cassette

A

GREEN

148
Q

BONE SAMPLE

Tissue cassette

A

WHITE

149
Q

SAMPLE IS SKIN

Tissue cassette

A

GRAY

150
Q

lymph node

Tissue cassette

A

PINK