[1S] UNIT 1.2 Instrumentation in Histopathology Flashcards

1
Q

Medical Technologists are also known as

A

Clinical Laboratory Scientists

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2
Q

Overview of Different Laboratory Procedures

A

Numbering
Fixation
Decalcification*
Dehydration
Clearing
Impregnation
Embedding
Blocking
Trimming
Sectioning
Staining
Mounting
Labelling

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3
Q

A procedure in which we assign a specific code/id for a particular specimen as the cassette containing the specimen is too small to include every specific detail

A

Numbering/Accessioning

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4
Q

T/F: Numbering may be a combination of numbers and letters

A

T

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5
Q

T/F: Site numbers (different areas of an organ) are not included if several sites are used

A

F; may also be included

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6
Q

The process of preserving the tissues and its cellular constituents

A

Fixation

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7
Q

● To prevent autolysis
● For the pathologists to read clearly and correctly to get an accurate diagnosis

A

Fixation

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8
Q

Fixation reagent

A

Formalin

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9
Q

The process of removing calcium ions to soften the tissue for easier cutting

A

Decalcification

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10
Q

T/F: All specimens undergo decalcification

A

F; not all

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11
Q

Decalcification reagent

A

Nitric Acid

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12
Q

The process of removing water or excess water

A

Dehydration

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13
Q

Why is water removed in a specimen?

A

Excess water is removed so that infiltration is performed properly

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14
Q

Dehydration reagent

A

Alcohol

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15
Q

● The process of removing alcohol
● Alcohol is removed to perform Impregnation or infiltration properly

A

Clearing

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16
Q

Clearing agent

A

Xylene

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17
Q

● The process of infiltrating the tissue with solution that will fill up the natural spaces to maintain the integrity of the specimen appearance
○ This provides rigid support for the tissues
○ To maintain the appearance and shape of the tissue microscopically

A

Impregnation / Infiltration

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18
Q

Impregnation reagent

A

Paraffin Wax

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19
Q

● The process of forming tissue block using a mold
● This is needed to perform cutting on a particular site

A

Embedding & Blocking

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20
Q

● The process of removing excess wax
● _________ __________ is created so that we can prepare a tissue ribbon properly

A

Trimming
Truncated pyramid

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21
Q

The process of cutting the tissue into micro thin slices of tissue

A

Sectioning / Cutting

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22
Q

● The process of coloring the tissues to clearly identify cells clearly, correctly, and accurately
● To differentiate cell from one another

A

Staining

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23
Q

● The process of securing the slide with a coverslip (permanent)
● To protect the stained slide
● Performed to protect the slide during handling and storage

A

Mounting

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24
Q

● The process of providing a particular label
● Assigned number is also used for this process

A

Labelling

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25
Q

How many steps are in tissue processing without decalcification?

A

12

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26
Q

Characterized by the transfer of tissues, contained within a basket, through a series of stationary reagents arranged inline or in a circular carousel plan

A

AUTOMATED TISSUE PROCESSOR: Tissue Transfer Processors (Carousel-Type)

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27
Q

TISSUE TRANSFER PROCESSORS (CAROUSEL-TYPE)
● ___ reagent positions & ___ wax positions
● Capacity = __________

A

● 9-10 and 2-3
● 30-110 cassettes

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28
Q

are the instruments where sample tissue is placed

A

Cassettes

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29
Q

Characterized by processing fluids pumped to and from a retort in which the tissues remain stationary

A

AUTOMATED TISSUE PROCESSOR: Fluid Transfer Processors

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30
Q

FLUID-TRANSFER PROCESSORS
● _____ reagent stations with temperatures adjustable between ______
● ___ paraffin wax stations with variable temperature settings between ______
● Capacity: _________

A

● 10-12 reagent stations with temperatures adjustable between 30-45°C
● 3-4 paraffin wax stations with variable temperature settings between 48-68°C
● Capacity: 100-300 cassettes

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31
Q

ADVANTAGES OF USING BOTH TISSUE-TRANSFER AND FLUID-TRANSFER

T/F: Flexibility of reagent selection

A

T

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32
Q

ADVANTAGES OF USING BOTH TISSUE-TRANSFER AND FLUID-TRANSFER

T/F: The conditions and schedule are designed to provide optimum processing even for the small batches of tissues

A

T

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33
Q

TISSUE PROCESSING

Used for making tissue blocks

A

Tissue Embedding Center

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34
Q

TISSUE PROCESSING

Advantage:
○ It has a refrigeration system
■ The cold platform in the equipment
■ To solidify the tissue
■ Paraffin is solidified for the tissue to be stable

A

Tissue Embedding Center

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35
Q

TISSUE PROCESSING: TISSUE EMBEDDING CENTER

Run the tissue embedding center for about _ hours to make sure paraffin wax is melted in the paraffin chamber

A

5

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36
Q

TISSUE PROCESSING: TISSUE EMBEDDING CENTER

T/F: Not all blocking steps can be performed with the help of this machine

A

F; All blocking steps can be performed

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37
Q

EMBEDDING PROCESS

What temperature should the paraffin be and wax melting point?

A

61°C
55-57°C (wax melting point)

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38
Q

EMBEDDING PROCESS

T/F: Place cassettes in the paraffin reservoir in numerical order

A

T

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39
Q

PARTS OF THE TISSUE EMBEDDING CENTER

The cool system of the Tissue Embedding Center

A

Freon Refrigeration System

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40
Q

PARTS OF THE TISSUE EMBEDDING CENTER

Where embedded tissues, with their respected molds to hasten solidification

A

Cold Orientation Platform

41
Q

PARTS OF THE TISSUE EMBEDDING CENTER

○ A button that provides a non clogging flow of molten paraffin for casting the mold
○ Same button just like in water dispenser

A

Microswitch Dispenser

42
Q

PARTS OF THE TISSUE EMBEDDING CENTER

Hot orientation platform materials

A

Forceps
Tissue
Metal Mold

43
Q

PARTS OF THE TISSUE EMBEDDING CENTER

T/F: During embedding, the orientation is provided by the pathologist

A

T

44
Q

PARTS OF THE TISSUE EMBEDDING CENTER

○ Top left part of the machine
○ Above the cold orientation platform
○ Catches excess paraffin

A

Waste Drawer

45
Q

PARTS OF THE TISSUE EMBEDDING CENTER

Area where we can preheat the forceps

A

Hot Well

46
Q

Basic instrument used that is capable of cutting section at a predetermined thickness by sliding the block into a cutting tool which is fixed and attached to the machine.

A

Microtomy

47
Q

PRINCIPLE: A spring balance teeth or pawl is brought into contact with, and turns a ratchet feed wheel connected to a micrometer screw, which in turn rotated, moving the tissue block at a predetermined distance towards the knife for cutting sections at uniform thickness.

A

Microtomy

48
Q

KINDS OF MICROTOME

● Invented by Paldwell Trefall in 1881
● Simplest among the different types of microtome

A

Rocking Microtome

49
Q

KINDS OF MICROTOME

● Consist of a heavy base and two arms:
● Used to cut small and large blocks of paraffin tissues.
● Not recommended for serial sections because tissues are cut in slightly curved planes

A

Rocking Microtome

50
Q

KINDS OF MICROTOME

● Disadvantage:
○ Restrictions in size of tissue block that can be cut
○ Difficulty of reorienting the block

A

Rocking Microtome

51
Q

KINDS OF MICROTOME

● Minot microtome
● Invented by Minot in 1885-1886

A

Rotary Microtome

52
Q

KINDS OF MICROTOME

Most common type used for both routine and research laboratories

A

Rotary Microtome

53
Q

KINDS OF MICROTOME

● Operated by the rotation of the flywheel
● Causes reciprocal motion of the knife over the block
● Thickness of the section being automatically regulated by the ratchet feed wheel

A

Rotary Microtome

54
Q

ROTARY MICROTOME DIFFERENCE FROM THE ROCKING MICROTOME

T/F: The knife and the block holder are brought together by upward and vertical motions

A

T

55
Q

ROTARY MICROTOME DIFFERENCE FROM THE ROCKING MICROTOME

T/F: Cuts sections in a perfectly flat plane

A

T

56
Q

ROTARY MICROTOME DIFFERENCE FROM THE ROCKING MICROTOME

● Heavier and more stable
● More complex in design and construction
● More expensive

A

T

57
Q

ROTARY MICROTOME DIFFERENCE FROM THE ROCKING MICROTOME

T/F: The blade is placed in a blade-down position which is relatively dangerous

A

F; blade-up

58
Q

KINDS OF MICROTOME

● Developed by Adams in 1789
● Especially recommended for cutting extremely hard and rough tissue blocks

A

Sliding Microtome

59
Q

KINDS OF MICROTOME

● The knife is moved horizontally against a fixed block which progresses against it in an inclined plane
● Most dangerous type of microtome

A

Sliding Microtome

60
Q

KINDS OF MICROTOME

Consist of two movable pillars holding the adjustable knife clamps, allowing the knife to be set at an angle for cutting celloidin sections

A

Base-Sledge Microtome

61
Q

KINDS OF MICROTOME

Favored in laboratories where very hard tissue or large blocks are usually sectioned

A

Base-Sledge Microtome

62
Q

KINDS OF MICROTOME

• Suited for sectioning specimens embedded in all forms of media
• Comparatively more stable

A

Base-Sledge Microtome

63
Q

KINDS OF MICROTOME

The block remains stationary while the knife is moved backward and forward during the process of sectioning.

A

Standard Sliding Microtome

64
Q

KINDS OF MICROTOME

• Developed mainly for cutting celloidin-embedded tissue blocks
• Inherently more dangerous

A

Standard Sliding Microtome

65
Q

KINDS OF MICROTOME

• Invented by Queckett in 1848
• The stage for block holder is hollow and perforated around its perimeter, attached to a reinforced flexible lead pipe thru which carbon dioxide passes from a cylinder.

A

Freezing Microtome

66
Q

KINDS OF MICROTOME

• A simple lever operated valve allows the release of rapid intermittent burst of carbon dioxide
• A second cooling device for lowering temperature of the knife to facilitate sectioning.

A

Freezing Microtome

67
Q

KINDS OF MICROTOME

• Used to cut undehydrated tissues in a frozen state
• When histological demonstration of fat is needed

A

Freezing Microtome

68
Q

KINDS OF MICROTOME

• When certain neurological structures are to be
studied
• When sensitive tissue constituents to be studied are damaged or destroyed by heat.

A

Freezing Microtome

69
Q

KINDS OF MICROTOME

• Consists of microtome kept inside a cold chamber which has been maintained at a temperature between -5 to -30°C (-20°C) by an adjustable thermostat
• Capable of freezing fresh tissues within 2-3 minutes
• Can cut sections of 4µ with ease.

A

FREEZING MICROTOME: Cryostat

70
Q

KINDS OF MICROTOME

Provides a means of preparing thin sections of fresh frozen tissues for fluorescent antibody staining techniques or histochemical enzyme studies.

A

FREEZING MICROTOME: Cryostat

71
Q

KINDS OF MICROTOME

Most commonly used for rapid preparation of urgent tissue biopsies for intraoperative diagnosis.

A

FREEZING MICROTOME: Cryostat

72
Q

are used to mount tissues in the Cryotome

A

OCTs

73
Q

KINDS OF MICROTOME

Primarily used for cutting tissue sections at 0.5 micra for electron microscopy

A

Ultrathin Microtome

74
Q

KINDS OF MICROTOME

The knife used for cutting the ultrathin sections consists mainly of selected fragments of ______ ______ ______.

A

broken plate glass

75
Q

KINDS OF MICROTOME: ULTRATHIN MICROTOME

The specimen used is small, fixed in ______ ________, and embedded in plastic.

A

osmium tetroxide

76
Q

Other Kinds of Microtome

A
  1. Vibrating Microtome
  2. Hand Microtome
  3. Saw Microtome
  4. Laser Microtome
  5. Computerized Microtome
77
Q

CARE OF THE MICROTOME

T/F: All the accumulated paraffin and small pieces of tissues must be brushed away with soft brush and not allowed to stay in the microtome

A

T

78
Q

CARE OF THE MICROTOME

T/F: After drying the machine and knife holder, the parts should be wiped with xylene

A

F; xylol not xylene

79
Q

CARE OF THE MICROTOME

T/F: Movable portions should be oiled thoroughly to prevent rusting

A

T

80
Q

CARE OF THE MICROTOME

T/F: Microtome must always be covered when not in use

A

T

81
Q

Trimming and section-cutting are done with a?

A

Microtome Knife

82
Q

TYPES OF MICROTOME KNIVES

• Usually 25 mm in length
• One side of the knife is flat while the other is concave.

A

Plane-Concave Knife

83
Q

TYPES OF MICROTOME KNIVES

• Usually 120 mm in length
• With both sides concave, recommended for cutting
paraffin embedded sections on a rotary microtome.

A

Biconcave Knife

84
Q

TYPES OF MICROTOME KNIVES

• Usually 100 mm in length
• Have both sides straight
• Recommended for frozen sections or for cutting extremely hard and tough specimens embedded in
paraffin blocks using a base-sledge type or sliding mircrotome.

A

Plane-Wedge Knife

85
Q

TYPES OF MICROTOME KNIVES

• Have a sharp cutting edge that can cut 2-4 µ thick sections with ease.
• Cheaper to use.

A

Disposable Blades

86
Q

TYPES OF MICROTOME KNIVES

• Used for trimming and semi-thin sectioning of tissue blocks for electron microscopy.
• Commercially prepared
• Should be prepared and stored in dust-free boxes with lids, just before use, to avoid contamination.

A

Glass Knives

87
Q

TYPES OF MICROTOME KNIVES

• Used to cut any type of resin block for electron microscopy
• Are brittle and expensive, but very durable

A

Diamond Knives

88
Q

TYPES OF MICROTOME KNIVES

They are already mounted in a metal block designed to fit directly into the knife holder of the ultrathin microtome when purchased.

A

Microtome Knives

89
Q

OTHER EQUIPMENT FOR TISSUE SECTIONING

• Temperature: About 10°C below the melting point of the paraffin wax.
• A small amount of detergent may be added to water to reduce surface tension and allow section to flatten out.

A

Floatation Water Bath

90
Q

OTHER EQUIPMENT FOR TISSUE SECTIONING

Temperature is set at the melting point of the wax, to 2-5 degrees Celsius above*

A

Drying Oven or Hot Plate

91
Q

OTHER EQUIPMENT FOR TISSUE SECTIONING

● To melt the paraffin
● Can be used to remove excess paraffin so that only the tissue is on the slide

A

Microwave

92
Q

OTHER EQUIPMENT FOR TISSUE SECTIONING

Both are needed for handling sections during cutting and removing folds and creases during “floating out” in waterbath.

A

Forceps & Squirrel/Camel Hairbrush

93
Q

Staining equipment should be?

A

Acid and alkali resistant, high temperature resistant

94
Q

OTHER EQUIPMENT

Programmed to perform H&E routine staining without any supervision. This has a continuous load and unload drawer access and robotic arm system that routinely transfers stained slides to another reagent or stain.

Recommended in a busy histopathology laboratory

A

Automated Stainer/Autostainer

95
Q

OTHER EQUIPMENT

These are covered glass vessels or containers, usually with grooves inside, and are used as containers of staining reagents during manual staining procedures.

A

Coplin Staining Jars

96
Q

OTHER EQUIPMENT

These are covered glass vessels or containers, usually with grooves inside, and are used as containers of staining reagents during manual staining procedures.

A

Coplin Staining Jars

97
Q

OTHER EQUIPMENT

Used to hasten the process of heating the slides after fishing the tissue section in the floatation water bath during sectioning in preparation for staining.

Up to 6 slide racks (30 slide capacities)

A

Slide Dryer

98
Q

PARTS OF TISSUE EMBEDDING CENTER

It filters particles or sediments

A

Microscreen

99
Q

PARTS OF TISSUE EMBEDDING CENTER

It forms a penthouse that contains molten paraffin. It has a microswitch dispenser usually on the lower left od the penthouse.

This chamber maintains an optimal paraffin temperature with adjustable thermostatic control

A

Paraffin Melting Chamber