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HISTOPATH LAB > Instrumentation in Histopathology > Flashcards

Instrumentation in Histopathology Flashcards

Based on PPT

1
Q

Order of tissue processing

A

(1) Numbering/Accessioning
(2) Fixation
(3) Decalcification
(4) Dehydration
(5) Clearing
(6) Impregnation
(7) Embedding
(8) Blocking
(9) Trimming
(10) Sectioning
(11) Staining
(12) Mounting
(13) Labelling

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2
Q

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

A

Tissue-transfer processors (carousel-type)

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3
Q

9-10 reagent positions and 2-3 wax positions
Capacity: 30-110 cassettes

A

Tissue-transfer processors (carousel-type)

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4
Q

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

A

Fluid-transfer processors

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5
Q

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

A

Fluid-transfer processors

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6
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

Microtome

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7
Q

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

A

Microtome Principle

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8
Q

Holds the tissue
Moves while the knife is stationary
Movement is usually forward then downwards
Controlled by spring balance teeth/pawl and the ratchet feed wheel connected to the micrometer screw that is calibrated to move so that the section would be as thin as prescribed by the protocol of the microtome

A

Chunk/Block/Tissue block

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9
Q

Specimen holder
Specimen block
Knife
(Sometimes included : Copper mesh Grid)
Ratchet Feed wheel
Adjustment screws

A

Essential Parts of microtome

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10
Q

Cuts the block at a predetermined thickness.
It produces tissue ribbons that are connected to each other

A

Knife

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11
Q

Line up the tissue block in the proper position
Adjust the thickness for the sections

A

Adjustment screws

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12
Q

Rocking microtome
Rotary microtome
Sliding microtome
Freezing microtome
Ultrathin microtome

A

Kinds of Microtome

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13
Q

Manually operated
Invented by Paldwell trefall in 1881
Simplest among the different types of microtome
Consists of a heavy base and two arms

A

Rocking Microtome

(Cambridge rocking microtome)

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14
Q

Name derived from the rocking action of the crossarm

A

Rocking microtome

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15
Q

Can cut up to 10-12µm tissue sections

A

Rocking microtome

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16
Q

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

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17
Q

Disadvantage of this microtome:
Restrictions in size of tissue block that can be cut
Difficulty of reorienting the block

A

Rocking Microtome

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18
Q

Manually operated
Minot microtome
Invented by Minot in 1885-1886
Most common type used for both routine and research laboratories

A

Rotary Microtome

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19
Q

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

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20
Q

Difference of Rotary microtome from Rocking microtome

A

The knife and the block holder are brought together by upward and vertical motions (cuts sections in a perfectly flat plane)
Heavier and more stable
More complex in design and construction
More expensive
The blade is placed in a blade-up position which is relatively dangerous

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21
Q

Manually operated
Discovered by Adams in 1789
Especially recommended for cutting extremely hard and rough tissue blocks
Most dangerous type of microtome

A

Sliding Microtome

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22
Q

Two kinds of sliding microtome

A

Base-sledge microtome
Standard sliding microtome

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23
Q

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

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24
Q

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

A

Base-sledge microtome

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25
Q

Suited for sectioning specimens embedded in all forms of media

A

Base-sledge microtome

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26
Q

Comparatively more stable

A

Base-sledge microtome

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27
Q

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

A

Standard sliding microtome

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28
Q

Developed mainly for cutting celloidin-embedded tissue blocks

A

Standard sliding microtome

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29
Q

Inherently more dangerous

A

Standard sliding microtome

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30
Q

Base-sledge microtome

Used in ALL FORMS of media
Block is ___
Knife is ____

A

Moving
Stationary

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31
Q

Common disadvantage of base-sledge microtome

A

slower than the rotary and rocking microtomes

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32
Q

Common tissues used in base-sledge microtome

A

Whole brain

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33
Q

Standard sliding microtome

Block is ___
Knife is ___

A

Stationary
Moving

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34
Q

Semi-automated
Invented by Queckett in 1848

A

Freezing microtome

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35
Q

The stage for the block holder is hollow and perforated around its perimeter, attached to a reinforced flexible lead pipe thru which carbon dioxide (freezing agent) passes from a cylinder

A

Freezing microtome

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36
Q

Designed for cutting frozen sections
No fixative is used prior to using this microtome

A

Freezing microtome

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37
Q

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

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38
Q

Used to cut undehydrated tissues in a frozen state

A

Freezing microtome

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39
Q

When histological demonstration of fat is needed

A

Freezing microtome

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40
Q

When certain neurological structures are to be studied

A

Freezing microtome

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41
Q

When sensitive tissue constituents to be studied are damaged or destroyed by heat

A

Freezing microtome

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42
Q

Kind of freezing microtome

A

Cyrostat

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43
Q

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

A

Cyrostat

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44
Q

Capable of freezing fresh tissues within 2-3 minutes

A

Cyrostat

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45
Q

Can cut sections of 4µ with ease

A

Cyrostat

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46
Q

Advantages of Cyrostat

A

Provides a means of preparing thin sections of fresh frozen tissues for fluorescent antibody staining techniques (immunohistochemistry)a or histochemical enzyme studies (microstructure and enzyme/antigen integrity)

47
Q

Most commonly used for rapid preparation of urgent tissue biopsises for intraoperative diagnosis

A

Cyrostat

48
Q

Used to mount tissues in the Cyrotome

A

OCTs

49
Q

Semi automated or automated
Primarily used for cutting tissue sections at 0.5 micra for electron microscopy

A

Ultrathin microtome

50
Q

The knife used for cutting the ultrathin sections consists mainly of selected fragments of broken plate glass

A

Ultrathin microtome

51
Q

The specimen used is small, fixed in osmium tetroxide and embedded in plastic

A

Ultrathin microtome

52
Q

Other kinds of microtome

A

Vibrating microtome
Hand microtome
Saw microtome
Laser microtome
Computerized microtome

53
Q

Also called as “vibratome”
Designed to cut sections from fresh and fixed materials from animal or botanical sources

A

Vibrating microtome

54
Q

Uses high-speed vibrations produced by safety razor blades which provide the cutting power, allowing the resultant cut to be made with less pressure than would the stationary blade be

A

Vibrating microtome

55
Q

Mostly used for rigid botanical materials
For the hard stems of plants

A

Hand microtome

56
Q

Designed to cut sections from very hard materials like undecalcified bone, teeth, glass and ceramics

A

Saw microtome

57
Q

Samples are usually embedded in resin and are moved extremely slowly against a diamond coated saw (slices through the sample) rotating at 600 rpm

A

Saw microtome

58
Q

Cannot produce very thin sections, the product is about 20 micra even using a diamond knife

A

Saw microtome

59
Q

Used for tissue that are embedded in one of the tougher media

A

Saw microtome

60
Q

Automatic microtome
Designed for non-contact sectioning inside biological tissues without causing thermal damage

A

Laser microtome

61
Q

Can produce tissue sections from 5-100µ (microns)

A

Laser microtome

62
Q

Operates using a cutting action with the used of infrared laser

A

Laser microtome

63
Q

The preparation of the sample through embedding, freezing, or chemical fixation is not required therefore minimizing the artifacts from the preparation methods in the tissue

A

Laser microtome

64
Q

Automatic microtome
Dual purpose microtome can produce:
Rapid freezing sections for RFS, Routine paraffin sections for routine histopathology lab tests

A

Computerized microtome

65
Q

Equipped with:
Thermostatic switch
semiconductor freezing
cyroscalpel
cyroplate

A

Computerized microtome

66
Q

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

A

Care of the microtome

67
Q

After drying the machine and knife holder, the parts should be wiped with ___

A

xylol

68
Q

Movable portions should be ___ thoroughly to prevent rusting

A

oiled

69
Q

Microtome must always be ___ when not in use

A

covered

70
Q

Trimming and section-cutting are done with a

A

microtome knife

71
Q

Used for hard sharpening
Done to remove gross nicks or irregularities
heel to toe direction
uses honing stones

A

Honing

72
Q

Done to remove burns
toe to heel direction
uses horse leather

A

Stropping

73
Q

Types of microtome knives

A

Plane-concave knife
Biconcave knife
Plane-wedge knife
Disposable blades
Glass knives
Diamond knives

74
Q

Usually 25mm in length
one side of the knife is flat while the other is concave

A

Plane-concave knife

75
Q

Flat side of the plane-concave knife is recommended for cutting _____ in a standard sliding microtome

A

Celloidin-embedded

76
Q

Concave side of a plane-concave knife is used to cut ___ sections in a base-sledge, rotary, or rocking microtome

A

paraffin

77
Q

Used cutting sections from a material that is too hard to cut with a biconcave knife (Could also be used for sapphire materials)

A

Plane-concave knife

78
Q

Usually 120mm in length
With both sides concave

A

Biconcave knife

79
Q

Recommended for cutting paraffin embedded sections on a rotary microtome (also used on rocking microtomes and carbon dioxide-freezing microtomes)

A

Biconcave knife

80
Q

Most recommended blade for routine cutting in microtomy

A

Biconcave knife

81
Q

Not suitable for relatively hard materials which cause the edge to vibrate and produce the phenomenon known as chattering that destroys the microanatomy of the tissue

A

Biconcave knife

82
Q

Usually 100mm in length
have both sides straight

A

Plane-wedge knife

83
Q

Recommended for frozen sections or for cutting extremely hard and tough specimens embedded in paraffin blocks using a base-sledge type or sliding microtome

A

Plane-wedge knife

84
Q

Also used for paraffin blocks being cut in the rotary microtome

A

Plane-wedge knife

85
Q

has more rigidity than biconcave knife and plane-concave knife

A

Plane-wedge knife

86
Q

preferred or placed in a base ledge type or in a sliding microtome type when cutting paraffin

A

Plane-wedge knife

87
Q

Knife angles

A

Wedge Angle
Bevel Angle
Clearance Angle
Cutting Angle
Rake angle

88
Q

Refer to the position of the tissue block in reference to the knife

A

Angle

89
Q

Flat side of the tissue block in reference to the bottom part of the knife would be known as the :

A

Clearange angle (0-15°)

90
Q

Best angle to be used

A

5°

91
Q

Angle of the edges in a wedge knife
Body of the knife or blade
Normally 15°

A

Wedge Angle

92
Q

Angle of the very tip blade, between cutting facets
Normally 27-32°

A

Bevel angle

93
Q

Angle of the knife itself in reference to the tissue block (27-32°)

A

Bevel angle

94
Q

Angle between the block face and the lower facet of the knife
Dependednt on the tilt or knife holder
Set between 3-8°

A

Clearance angle

95
Q

When the clearance angle is too wide

A

The tip of the blade will scrape the block and chatter will result

96
Q

When the clearance angle is too small

A

the body of the blade will scrape the block and skipped sections or poor ribboning will result

97
Q

Angle between the block face and upper facet of the knife
This is not an angle that can be adjusted on a microtome

A

Cutting angle

98
Q

It is the result of the clearance angle and the upper bevel angle of the knife or blade

A

Cutting angle

99
Q

90° minus the angle of the upper facet of the knife
Subtract all the angles that have been set to 90°

A

Rake angle

100
Q

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

A

Disposable blades

101
Q

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

102
Q

Used to cut any type of resin block for electron microscopy

Brittle and expensive but very durable

They are already mounted in a metal bock designed to fit directly into the knife holder of the ultra-thin microtome when purchased

A

Diamond knives

103
Q

Other equipment for tissue sectioning

A

Floatation water bath
Drying oven or hot plate
Microwave
Forceps and squirrel/camel hairbrush
Clean slides and coverslips

104
Q

Floatation water bath

Temperature : About __ below the melting point of the paraffin wax

A

10°

105
Q

Floatation water bath

A small amount of detergent may be added to water to ___ and allow ____

A

Reduce surface tension

allow section to flatten out

106
Q

Floatation water bath

Capacity : ___ liters
__ inches (diameter) x __ inches (height)

A

2 liters
11 inches (diameter) x 4 inches (height)

107
Q

mostly used after microtomy to remove the wrinkles and crumpled areas of tissue ribbon

A

Floatation water bath

108
Q

Temperature is set at the melting point of the wax to 2-5°C above* (*Gregorios)

A

Drying oven or hot plate

109
Q

For rapid drying of tissue slides
The unstained tissue slides should be put here
For delicate tissues, a lower drying temperature must be used to avoid cracking and splitting of the sections

A

Drying oven or hot plate

110
Q

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

A

Microwave

111
Q

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

Both could be used for sectioning

A

Forceps and Squirrel/Camel Hairbrush

112
Q

Mostly needed for embedding

A

Forceps

113
Q

Used to fix sections in place

A

Forceps and Squirrel/Camel Hairbrush

HISTOPATH LAB (2 decks)

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