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Histopathology Lec&Lab > MODULE 4 - Sectioning > Flashcards

MODULE 4 - Sectioning Flashcards

1
Q

Involves cutting of tissues into thin slices

A

Sectioning

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2
Q

Size of paraffin sections

A

4-6 um

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3
Q

Size of celloidin sections

A

10-15 um

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4
Q

Size of frozen sections

A

4 um

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5
Q

Disadvantage of using cambridge/rocking microtome

A

Difficult in re-orienting tissues and restriction in tissue size of blocks

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6
Q

Cambridge/rocking microtome is made by

A

Trekfall

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7
Q

Most simple microtome

A

Cambridge/Rocking

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8
Q

Cambridge/Rocking microtome is made for

A

serial sections of large paraffin blocks

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9
Q

Thickness produced when using cambridge/rocking microtome?

A

10-12 micra

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10
Q

Routinely and most common microtome

A

Rotary

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11
Q

Rotary microtome is made by

A

Minot

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12
Q

Thickness produced when using rotary microtome

A

4-6 micra

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13
Q

Rotary microtome is used for

A

paraffin embedded tissues

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14
Q

Sliding microtome is made by

A

Adams

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15
Q

The most dangerous microtome and for what reason

A

Sliding microtome because of exposed knife

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16
Q

Example of wax used in sliding microtome

A

Ester wax

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17
Q

Sliding microtome is used for

A

celloidin embedded tissues and extremely hard tissues

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18
Q

Thickness produced when using sliding microtome

A

7-9 micra

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19
Q

2 types of sliding microtome

A
  1. Base sledge

2. Standard sliding

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20
Q

Most dangerous type of sliding microtome

A

Standard microtome

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21
Q

Freezing/cold knife uses

A

intermittent burst of CO2 to immediately harden block holder and tissues without prior fixations.

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22
Q

Freezing/cold knife has a

A

second cooling device to lower temp of knife

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23
Q

Freezing/cold knife is used for

A

frozen sections and heat-sensitive structures

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24
Q

Freezing/cold knife is used for demonstration of

A

fats and neurological structures

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25
Q

Freezing/cold microtome was invented by

A

Queckette

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26
Q

Dew line is

A

point of sections in which it can be cut at 10um

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27
Q

thickness and tissue block produced using freezing/cold knife microtome

A

thickness: 10-15 um
block: 3-5 mm

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28
Q

Type of microtome enclosed in a chamber and is primarily composed of rotary microtome

A

Cryostat/cold microtome

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29
Q

Cryostat/cold microtome is used for

A

fresh frozen tissues for fluorescent antibody staining and histochemical enzyme studies

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30
Q

Temperature used in cryostat/cold microtome

A

-5 to -30 degC

average -20 degC

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31
Q

Microtome used when tissues are usually embedded in plastic

A

ultrathin microtome

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32
Q

ultrathin microtome is used for

A

EM

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33
Q

thickness of tissues when using ultrathin microtome

A

0.5 um

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34
Q

process of removing excess paraffin wax from tissues once properly
fixed on the slide

A

DEPARAFFINIZATION

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35
Q

METHODS of DEPARAFFINIZATION

A
  1. Passing of slide over alcohol lamp
  2. Immersing slide in xylene
  3. Putting it on oven (55-60 degC)
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36
Q

Microtome knives that is 25mm long

A

Plane concave

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37
Q

Microtome knives that is 120mm long

A

Biconcave

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38
Q

Microtome knives that is 100mm long

A

Plane wedge

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39
Q

Plane concave flat side is used for?

concave side is for?

A

Flat- celloidin

Concave- paraffin

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40
Q

Biconcave is used for

A

paraffin embedded and rotary

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41
Q

Plane wedge is for

A

frozen sections and extremely hard tissues

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42
Q

Bevel angle is

A

angle formed between the cutting edges 27-32 deg

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43
Q

Clearance angle is also known as

A

Tilting/inclination angle

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44
Q

Clearance angle is

A

angle formed between surface of blocks and cutting edge of knife @0-15 deg but in fixation it is 5-10 deg to avoid uneven sections (alternate thin and thick)

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45
Q

In Clearance angle >10 indicates and <10 indicates

A

> 10 - low thickness

<10 - high thickness

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46
Q

Wedge angle is

A

angle formed in sides of wedge knife 14-15 deg

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47
Q

Thickness produced by disposable blades

A

2-4 um

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48
Q

Glass and diamond knives are used for

A

EM

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49
Q

Purpose of honing

A

to remove nicks

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50
Q

Honing uses what movement

A

heel to toe movement, edge first 20-30 times

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51
Q

Honing uses

A

hones/oil stones

lubricant

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52
Q

Oil stones used in honing

A

Belgium yellow
Arkansas
Fine carborundum

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53
Q

Lubricants used in honing

A 
soapy water
mineral oil
clove oil
xylene
liquid paraffin
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54
Q

Belgium yellow

A

gives best result

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55
Q

Arkansas

A

gives more polishing effect than belgium yellow

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56
Q

fine carborundum

A

for badly nicked knives, has coarser bite

57
Q

Alternatives for oil stones

A

Glassplates and diamantine

58
Q

What composes glass plates

A

powdered aluminum oxide with water = paste = ABRASIVE

59
Q

Diamantine is

A

for final polishing

60
Q

Polishing and sharpening of knife

A

stropping

61
Q

purpose of stropping

A

to remove burrss and other deformities formed by honing

62
Q

stropping is done with what movement and how many strokes

A

toe to heel, edge last 40-120 double strokes

63
Q

Strapping uses _____ made of _______

A

paddle strop, horse leather

64
Q

Strops are treated with

A

oil

65
Q

In stropping do not use ______ because it can ________

A

lubricant, mineral oil - blister and damage leather

66
Q

Purpose of floatation water bath

A

thermostatically controlled bath to remove folds and wrinkles to flatten ribbons

67
Q

Temperature of floatation water bath when in use

A

6-10 degC lower than wax melting point

specifically, 45-50 degC

68
Q

Product of section

A

thin slices , ribbons

69
Q

what is fishing out

A

removal of ribbon in float out bath

70
Q

3 methods for Drying of slides

A
  1. leave in incubator @37 degC overnight
  2. put in oven @50-60 degC for 2 hours
  3. in a hot plate @45-55 degC for 30-45 mins
71
Q

This is used to promote attachment to slides and prevent detachment

A

adhesives

72
Q

Routine tissue adhesive

A

Mayer’s egg albumin

73
Q

Mayer’s egg albumin is composed of

A

Egg white + glycerin + thymol crystals

74
Q

Thymol crystals is used to

A

prevent growth of molds

75
Q

Poly-L-Lysine is used for

A

immunohistochemistry

76
Q

APES (3-aminopropylthriethoxysilane) is used for

A

cytology

77
Q

With strong adhesive property

A

sodium silicate

78
Q

Adhesive that is a commercial syrup

A

sodium silicate

79
Q

Enumerate methods of freezing and their details

A 
Liquid nitrogen- most rapid, for histochem
Isopentane- liquid at RT
Aerosol sprays (cryokwik)
CO2- freezing microtome
Freon 2.2- high thermal conductivity
80
Q

Microtome parts and their details

A
  1. Block holder/chuck- where tissues are held in position
  2. Knife carrier/knife- disposable knives, for actual cutting
  3. Rotating/flying wheel- done mechanically to start the cutting process
  4. Pawl, ratched feed wheel, adjustment screw- lines up tissues in proper positions with knife
81
Q

A microtome that is theoretically not recommended for serial sections since tissues are cut in slightly curved planes.

A

Cambridge/rocking microtome

82
Q

Typically, sections are cut between ___________ µm using paraffin wax for diagnostic histology

A

3 and 5 um

83
Q

It was originally designed for cutting sections of very large blocks (whole brain).

A

Sliding microtome

84
Q

Microtomes used for serial sections (book)

A

Rotary

Sliding- basic sledge

85
Q

It is used to cut undehydrated thin to semi-thin sections of fresh, frozen
tissues

A

Freezing/cold knife

86
Q

capable of freezing fresh tissues within 2-3 minutes, and cutting sections of 4 µ with ease.

A

The Cryostat or Cold Microtome

87
Q

It is most commonly used for rapid preparation of urgent tissue biopsies for intraoperative diagnosis.

A

The Cryostat or Cold Microtome

88
Q

It is often housed in the frozen section room close to the operating room to allow direct consultation between surgeon and pathologist.

A

The Cryostat or Cold Microtome

89
Q

microtome equipped with a glass or gem grade diamond knife is used to cut very thin sections (typically 60 to 100 nanometer) of tissue embedded in epoxy resin.

A

Ultrathin microtome

90
Q

Thin sectioning for the TEM is often done with a

A

gem quality diamond knife

91
Q

After sectioning, all the accumulated paraffin and small pieces of tissues
must be brushed away with a soft brush and not allowed to stay in the
microtome, since this may later interfere with the cutting of tissue blocks.
TRUE OR FALSE

A

True

92
Q

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

A

xylol

93
Q

Prolonged and continuous application of the painted parts with xylene will

A

remove the paint

94
Q

Movable portions should not be oiled to prevent rusting

TRUE OR FALSE

A

False

95
Q

The microtome must always be covered when not in use

TRUE OR FALSE

A

True

96
Q

The microtome should be placed on an unstable bench, away from air drafts, doorways and passing staff.
TRUE OR FALSE

A

False

97
Q

Any air movement from air conditioners or other causes will not affect section handling
TRUE OR FALSE

A

False

98
Q

No fluid must enter the inside of the instrument during cleaning.
TRUE OR FALSE

A

True

99
Q

Have the instrument inspected at least __________ by a qualified service technician.

A

once a year

100
Q

Microtome that uses plane concave

A

rotary or rocking microtome

101
Q

Each knife should have its own corresponding back which should be interchanged with another, to keep the bevel angle.
TRUE OR FALSE

A

False

102
Q

Too soft cutting edges are likely to

A

dull easily

103
Q

too hard edges are likely to produce

A

nicks or jagged edges and irregularities on the knife edge

104
Q

A good cutting edge must be able to cut good sections from a paraffin wax block about

A

2-3 microns thick

105
Q

may be used for partially calcified materials, paraffin and frozen sections.

A

Safety razor blades

106
Q

Theoretically, the perfect and optimum cutting angle is obtained when the sides of the wedge knife are inclined at an angle of about

A

15 deg

107
Q

The degree of sharpness is __________ to the fineness of the abrasive used in sharpening

A

proportional

108
Q

Hones for manual sharpening when cutting edge has been rendered blunt or nicked

A

belgium yellow

109
Q

The surface of the hone is wiped clean with a soft cloth moistened with ______ in order to remove the scattered small particles of stones and metal.

A

xylene

110
Q

In the case of the Minot or plane-wedge knife, the knife is turned over so as to sharpen the other surface every

A

I0-20 strokes

111
Q

Speed in stropping should be avoided

True or False

A

True

112
Q

Leather strops are usually dry and require oiling before they are used.
Strops are usually treated with

A

vegetable oil (castor)

113
Q

The strop should not be used for at least ________ after treatment.

A

24-48 hrs

114
Q

generally used for trimming and semi-thin sectioning of tissue blocks for electron microscopy.

A

Glass Knives

115
Q

used to cut any type of resin block for electron microscopy.

A

diamond knives

116
Q

used to cut any type of resin block for electron microscopy.

A

Diamond Knives

117
Q

These tools are needed for handling sections during cutting, and for removing folds and creases on the sections during “floating out” in water bath.

A

Forceps (fine pointed or curved) and squirrel hair brush

118
Q

The quality of sections cut on a microtome suffer badly from these several (avoidable) causes.

A

Fecal material and hair

119
Q

Coarse facing is done on the microtome at approximately

A

30 microns at a time

120
Q

Cooling both the tissue and the wax will give them a similar consistency, and make sectioning easier
TRUE or FALSE

A

true

121
Q

In fine trimming, the knife is usually tilted at

A

0-15 deg

122
Q

provides better support for the harder elements in a specimen allowing thinner sections to be obtained.

A

cold wax

123
Q

Generally a fast, uniform cutting stroke produces the best results and the least compression.
TRUE or FALSE

A

False

124
Q

Do not stop and restart during a cutting stroke as this will produce bands of different thickness across the section.
TRUE OR FALSE

A

True

125
Q

Sections should not be left on the water bath for a long time (30 seconds will be enough)

A

to avoid undue expansion and distortion of tissue.

126
Q

When flattening out sections, 20% alcohol will produce

A

convection current

127
Q

expand the section to its original dimensions and ensure

that it is completely flat.

A

floatation

128
Q

not recommended because they can cause overheating and there is a risk of dust falling onto the section during the drying period.

A

hot plate

129
Q

Excessive heat can cause

A

boiling of water droplets underneath the section which will cause damage

130
Q

Dry sections for between

A

5 and 30 minutes

131
Q

Staining of serial sections should never be attempted unless they are completely dried.
TRUE OR FALSE

A

True

132
Q

charged slides or an adhesive must be used in

A 
antigen retrieval (IHC)
enzyme pretreatment (ISH)
prolonged incubation steps
133
Q

Extended storage (usually more than 3 days) of unstained formalin-fixed paraffin embedded slides will cause

A

loss of antigens

134
Q

When cutting sections for DNA or RNA extraction, all instruments and equipment must be pre-cleaned and wiped down with

A

RNAse

135
Q

type of water used for floating sections for RNA extraction.

A

molecular grade water

136
Q

Hardening of the celloidin block may be hastened by placing

A

chloroform

137
Q

To avoid dehydration and shrinkage in section cutting, what should be done?

A

lubricated with 60-70% ethanol

138
Q

essential for methods that require exposure of sections to acids and alkalis (especially ammoniacal silver solutions) during staining

A

adhesives

139
Q

One disadvantage of using albumin

A

retention of stain which causes dirty background

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