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HISTOPATHOLOGY > PREFINALS: TRIMMING AND MICROTOMY > Flashcards

PREFINALS: TRIMMING AND MICROTOMY Flashcards

1
Q

Removal of excess wax using knife or cutter after the wax block is removed from the tissue cassette or paper boat

A

TRIMMING

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2
Q

Formation of truncated pyramid and exposure of the tissue surface for ease of sectioning

A

TRIMMING

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3
Q

Allow tissue blocks to fit into the block holder of microtome

A

TRIMMING

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4
Q

At least 2mm should surround the tissue block

A

TRIMMING

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5
Q

TRIMMING 2 TYPES

A

o Coarse Trimming
o Fine Trimming

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6
Q

AKA Sectioning

A

MICROTOMY

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7
Q

Formation of uniformly thin slices/sections/ribbons from the tissue block with the use of a microtome in order to facilitate studies under the microscope

A

MICROTOMY

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8
Q

Sections usually form ribbons due to slight heat generated between the block and the knife edge during the process of cutting

A

MICROTOMY

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9
Q

Complete ribbons are picked up with camel hair brush, forceps or fingers.

A

MICROTOMY

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10
Q

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

A

Microtome

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11
Q

Microtome Basic Parts:

A

 Block holder/chuck/cassette clamp
 Knife Carrier and Knife
 Pawl, Ratchet Feed Wheel and Adjustment Screws

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12
Q

where the tissue is held in
position

A

Block holder/chuck/cassette clamp

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13
Q

for actual cutting of tissue sections

A

Knife Carrier and Knife

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14
Q

to line up the tissue block in proper position with the knife, and to adjust the proper thickness of the tissue

A

Pawl, Ratchet Feed Wheel and Adjustment Screws

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15
Q

Brush away accumulated paraffin and small pieces of tissues with _______ after sectioning

A

soft brush

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16
Q

Excess paraffin and tissues may later on ______ with the cutting of tissue blocks

A

interfere

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17
Q

may also be used for cleaning some parts of the microtome

A

Xylene

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18
Q

_____ to prevent rusting.

A

Oil movable parts

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19
Q

______ to prevent accumulation of dust.

A

Cover microtome

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20
Q

The cutting rate depend upon the type of tissue, the size of the block and the model of the microtome that is used

A

TRUE

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21
Q

Rocking or Cambridge
Inventor:

A

Paldwell Trefall

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22
Q

Simplest

A

Rocking or Cambridge

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23
Q

Rocking or Cambridge
Cutting section:

A

10-12µm

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24
Q

For small and large paraffin blocks

A

Rocking or Cambridge

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25
Q

not for serial sections because of slightly curved planes

A

Rocking or Cambridge

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26
Q

Rotary Inventor:

A

Minot

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27
Q

Most common

A

Rotary

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28
Q

Rotary Media:

A

Paraffin

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29
Q

Excellent for serial sections

A

Rotary

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30
Q

Sliding Inventor:

A

Adams

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31
Q

Sliding Media:

A

Celloidin

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32
Q

Very hard and rough tissue blocks

A

Sliding

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33
Q

Sliding Types:

A

o Base-sledge
o Standard sliding

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34
Q

more dangerous d/t moving knife

A

Standard sliding

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35
Q

Freezing Inventor:

A

Queckett

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36
Q

Frozen sections (undehydrated tissues like fat) for rapid diagnosis

A

Freezing

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37
Q

Stage for block holder is hollow and perforated, attached to a flexible lead pipe containing carbon dioxide.

A

Freezing

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38
Q

CO2 as propellant/freezing agent (2 to 3 minutes)

A

Freezing

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39
Q
A

Cryostat/Cold Microtome

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40
Q

More common than freezing microtome

A

Cryostat/Cold Microtome

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41
Q

STAT frozen section (intraoperative diagnosis)

A

Cryostat/Cold Microtome

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42
Q

Chamber maintained -5 to -30OC with rotary microtome inside

A

Cryostat/Cold Microtome

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43
Q

Cryostat/Cold Microtome Cutting Section:

A

4 µm

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44
Q

Microtome Knives:

A
  • Plane-Concave
  • Biconcave
  • Plane-Wedge
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45
Q

Length

Plane-Concave

A

25mm

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46
Q

Length

Biconcave

A

120mm

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47
Q

Length

Plane-Wedge

A

100mm

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48
Q

Characteristic

Plane-Concave

A
  • One side is flat
  • Other side is concave
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49
Q

Characteristic

Biconcave

A

Both sides concave

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50
Q

Characteristic

Plane-Wedge

A

Both sides straight

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51
Q

Embedding Medium

Plane-Concave

A
  • Celloidin
  • Paraffin
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52
Q

Embedding Medium

Plane-Concave:
Less concave

A

Celloidin

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53
Q

Embedding Medium

Plane-Concave:
More concave

A

Paraffin

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54
Q

Embedding Medium

Biconcave

A

Paraffin

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55
Q

Embedding Medium

Plane-Wedge

A

Paraffin

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56
Q

Microtome

Plane-Concave

A
  • Sliding
  • Base-sledge, Rotary or Rocking
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57
Q

Microtome

Plane-Concave:
Celloidin

A

Sliding

58
Q

Microtome

Plane-Concave:
Paraffin

A

Base-sledge, Rotary or Rocking

59
Q

Microtome

Biconcave

A

Rotary

60
Q

Microtome

Plane-Wedge

A

Base-sledge

61
Q

A good cutting edge must be able to cut good sections from a paraffin wax block about ____, without any serration

A

2-3microns thick

62
Q

likely to produce nicks or jagged edges

A

Too hard cutting edges

62
Q

likely to become easily dull

A

Too soft cutting edges

63
Q

Angles:

A
  1. Bevel Angle: 27° to 32°
  2. Cutting angle (15°)
  3. Clearing Angle
64
Q

Angle between the cutting edges

A

Bevel Angle: 27° to 32°

65
Q

Maintained by slide-on back (spring-loaded, semi-circular metal sheet slipped onto the knife)

A

Bevel Angle: 27° to 32°

66
Q

angle between the face of a cutting tool and the surface of the work

A

Cutting angle (15°)

67
Q
  • knife should be inclined at 5-10° from the cutting plane so that the cutting facets (bevel angle) will not compress the block during the process of cutting.
A

Clearing Angle

68
Q

Other Knives and Blades:

A
  1. Disposable Blades
  2. Glass Knives/Ralph Knives
  3. Diamond knives
  4. Safety razor blades
69
Q

Widely used now because cheaper; honing and stropping are no longer common practice;

A

Disposable Blades

70
Q

coated with polytetrafluoroethylene (for ease of ribboning)

A

Disposable Blades

71
Q

E.g. Magnetic knives in cryostat

A

Disposable Blades

72
Q

For ultrathin microtomes

A

Glass Knives/Ralph Knives

73
Q

For resin blocks on ultrathin microtomes; brittle and expensive

A

Diamond knives

74
Q

Easily replaced when dull and produces good tissue sections same as with microtome knives

A

Safety razor blades

75
Q

For partially calcified materials, paraffin and frozen sections.

A

Safety razor blades

76
Q

Unsatisfactory for sections less than 10micra

A

Safety razor blades

77
Q

Badly nicked knives with blunted ends have to undergo sharpening in order to ensure optimum sectioning of tissue blocks

A

SHARPENING

78
Q

Sharpening of the knives involves 2 stages, namely:

A

1) Honing
2) Stropping

79
Q

Removal of nicks and irregularities on the knife edges

A

Honing

80
Q

Natural sharpening stone or hard grinding surface

A

Hones

81
Q

Wide enough to sufficiently support and prevent the rocking of the knife

A

Hones

82
Q

Long enough to allow the whole length of knife edge to be sharpened in a single stroke

A

Hones

83
Q

TYPES OF Hones:

A
  • Belgium Yellow
  • Arkansas
  • Fine Carborundum
  • Plate Glass (8x3x1in)
84
Q

most common; best result

A

Belgium Yellow

85
Q

has more polishing effect

A

Arkansas

86
Q

coarser; for badly nicked knives

A

Fine Carborundum

87
Q

excellent

A

Plate Glass (8x3x1in)

88
Q

Lubricants

A

o Soapy water
o Mineral oil
o Clove oil
o Xylene
o Liquid paraffin

89
Q

May be used for grinding and removing nicks

A

Flat Glass plate with finely powdered aluminum oxide

90
Q

For final polishing

A

Diamantine

91
Q

Procedure: Heel to Toe movement, Edge first

A

Honing

92
Q

Procedure: Heel to Toe movement, Edge first:

A
  1. Clean hone with xylene to remove scattered particles of stones and metal
  2. Cover with lubricant
  3. Knife is fitted to its corresponding back, placed on one end of the hone with cutting edge first
  4. With cutting knife edge first, the “heel” (handle end) is drawn obliquely or diagonally towards the operator on the stone until the “toe” (head portion) is reached.
  5. Honing is then continued until all the teeth in the knife edge have been eradicated”
  6. Washed with water after using to simply remove the metal collected during the process
  7. After honing, wipe off the oil or soap from the knife with xylene, then strop it thoroughly……
93
Q

_____ per surface for Minot or Plane-wedge knife

A

10-20 strokes

94
Q

For plane-concave knives,_____

A

only the concave surface should be rubbed on the hone

95
Q

Plane-wedge & plane-concave is provided with “backs” to maintain the ____ throughout honing

A

correct bevel angle (27-32°)

96
Q

Utilizes a machine that make use of vibrating frosted slide plate or wheel driven by electrical motor.

A

Mechanical Honing

97
Q

The knife is pressed against the flat side of a rotating glass wheel.

A

Mechanical Honing

98
Q

30 double strokes - given to each side of the knife.

A

Mechanical Honing

99
Q

Advantages: Time-saving; produce well sharpened knives with uniform bevels

A

Mechanical Honing

100
Q

Disadvantage: Expensive

A

Mechanical Honing

101
Q

Removal of burrs and polishing of cutting edge

A

Stropping

102
Q

Stropping Materials:

A

Paddle strop made of horse leather attached to a solid back, in order to prevent sagging

103
Q

Usually dry thus require oiling

A

Paddle strop made of horse leather attached to a solid back, in order to prevent sagging

104
Q

Vegetable oil (e.g. castor oil) applied on the back of the horse leather

A

Paddle strop made of horse leather attached to a solid back, in order to prevent sagging

105
Q

Not mineral oil because it tends to blister and destroy the leather

A

Paddle strop made of horse leather attached to a solid back, in order to prevent sagging

106
Q

Procedure: Toe to Heel movement, Edge Las

A

Stropping

107
Q

The procedure is the reverse of honing

A

Toe to Heel movement, Edge Las

108
Q

Procedure: Toe to Heel movement, Edge Last:

A
  1. The knife is fitted with its appropriate knife back
  2. Knife is laid obliquely on the strop and with the cutting edge behind
  3. Edge last is pushed backward and drawn forward
109
Q

Procedure: Toe to Heel movement, Edge Last
Precautions:

A

 The knife should always be wiped clean with a “soft” cloth before and after series of stropping (NEVER use paper or cloth)
 The knife edge is the oiled or greased to preventing it from rusting
 Pressure during the first stropping strokes should be quite light since the natural compressibility of the leather is what actually does the work
 Speed in stropping should be avoided
 Wax must not be allowed to come in contact with the strop.

110
Q

40-120 double strokes:

A
  • Plane wedge knife
  • Plane – Concave knife
111
Q

both sides are required for stropping

A

Plane wedge knife

112
Q

only the concave surface should be stropped

A

Plane – Concave knife

113
Q

TYPES OF TISSUE SECTIONS:

A
  1. Paraffin
  2. Celloidin
  3. Frozen Sections
114
Q

4-6µm

A

Paraffin

115
Q

Successive sections will usually stick edge-to-edge (knife) due to local pressure with each cutting stroke, thereby forming a ribbon. (remedy: cut slowly)

A

Paraffin

116
Q

Sections are removed in ribbons of ten to allow easy location of serial sections.

A

Paraffin

117
Q

The blocks are trimmed in the same manner as in paraffin blocks

A

Celloidin

118
Q

To avoid dehydration and shrinkage, sections are usually cut by the wet method, both the sections and the block being kept moist with 70% alcohol during cutting.

A

Celloidin

119
Q

Celloidin sections do not come off in ribbons and have to be collected into 70% alcohol immediately

A

Celloidin

120
Q

Methods of preparing frozen section

A

Frozen Sections

121
Q

Frozen Sections types:

A

1) Cold knife procedure
2) Cryostat procedure (cold microtome)

121
Q

10-15µm

A

Celloidin

122
Q

Sections are floated out on a water bath set at 45-50°(approx. 6-10°C lower than the melting point of the wax used for embedding the tissue.)

A

FLOTATION/FLOATING-OUT

123
Q

For paraffin sections

A

FLOTATION/FLOATING-OUT

124
Q

5 to 10OC↓MP of Wax

A

Flotation Bath

125
Q

Inside is specifically colored enamel black

A

Flotation Bath

126
Q

TSEs flatted after 30sec; removes tse wrinkling

A

Flotation Bath

127
Q

Flotation Bath
Dimensions:

A

d=11in, h=4in, 2L capacity

128
Q

Slides SIZE:

A

76x25mm, 1-1.2mm thick, frosted)

128
Q

Regulated temp. to flatten the sections and prepare them for mounting into the slides/slider

A

T

129
Q

Sections should not be left on the water bath for a long time (30 seconds will be enough) to avoid undue expansion and distortion of the tissue

A

T

130
Q

___may be used for adhesion of tissue to the slide

A

ADHESIVES

131
Q

may be removed by stretching the
sections gently

A

Folds or creases sections

132
Q

may be teased out beneath the sections by means of needle

A

Bubbles

133
Q

Selected sections for staining should be fished out in a vertical position

A

FLOTATION/FLOATING-OUT

134
Q

Mounted sections are placed in a paraffin oven to dry

A

DRYING THE SLIDES

135
Q

45 – 55°c for:

A
  • enzyme digestion
  • chemical extraction
  • metallic impregnation
  • enzyme localization technique
136
Q

Hot plates are not recommended because they can cause:

A

 Overheating
 Dust falling – onto the section during drying period

137
Q

Metal racks with 25-slide divisions are used to store the mounted sections during the drying process which usually takes 5 minutes in the heated oven. Once dry, the whole rack of slides can be taken for manual staining.

A

DRYING THE SLIDES

HISTOPATHOLOGY (13 decks)

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