MICROTOMY Flashcards

1
Q

previously processed tissue / tissue block is trimmed and cut into uniformly thin slices or sections

A

Microtomy

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2
Q

principle of microtomy

A

a spring balanced teeth 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

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3
Q

when was rocking (cambridge) microtome invented

A

1881

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4
Q

who invented the rocking (cambridge) microtome

A

Paldwell Trefall

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5
Q
  • the simplest type of microtome for small and large paraffin-embedded blocks
A

rocking microtome

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6
Q

thickness of rocking microtome

A

10-12u

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7
Q

one of its disadvantage is, it will not form serial sections

A

rocking microtome

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8
Q

part of the rocking microtome that carries the block holder

A

upper arm

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9
Q

part of the rocking microtome that supports the column

A

lower arm

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10
Q

commonly used microtome for paraffin-embedded sections

A

rotary microtome

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11
Q

allows production of excellent serial sections

A

rotary

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12
Q

heavier and more stable microtome with up and down vertical movement which results to perfectly flat planes

A

rotary

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13
Q

who invented the rotary m

A

minot

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14
Q

when was the rotary microtome invented

A

1885-86

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15
Q

thickness of rotary m

A

4-6u

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16
Q

2 types of sliding m

A
  1. standard sliding
  2. base-sledge
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17
Q

the thickness of sliding m

A

4-9u

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18
Q

who invented the sliding m

A

Adams

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19
Q

when was the sliding m invented?

A

1789

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20
Q

sliding m that consists of 2 movable pillars holding the adjustment knife clamps for hard tissues and large blocks

A

base sledge

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21
Q

sliding m that is prone to danger

A

standard sliding

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22
Q

block remains stationary, the microtome moves in a forward and back motion

A

standard sliding

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23
Q

for hard and tough tissue blocks in all forms of media

A

base-sledge

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24
Q

for electrom microscope

A

ultra thin microtome

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25
Q

thickness of ultra-thin m

A

0.5u

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26
Q

uses fragments of broken plate glass

A

ultrathin microtome

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27
Q

who invented the freezing microtome

A

Queckett

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28
Q

when was the freezing microtome invented

A

1848

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29
Q

for undehydrated tissues in frozen state

A

freezing microtome

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30
Q

for rapid diagnosis and for fats and tissue constituents that may be damaged

A

freezing microtome

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31
Q

thickness of freezing m

A

10-15u

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32
Q

source of cold temp for freezing microtome

A

carbon dioxide

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33
Q

sections to be focused under compound microtome

A
  1. plane-concave
  2. plane-wedge
  3. biconcave
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34
Q

for paraffin and celloidin embedded sections

A

plane-concave

35
Q

length of plane-concave

A

25mm

36
Q

for celloidin-embedded sections and for extremely hard tissues

A

plane-wedge

37
Q

length of plane-wedge knife

A

100mm

38
Q

for rotary microtome and for paraffin-embedded sections

A

biconcave

39
Q

Knives: sections to be focused under EM

A
  1. Diamond-edge
  2. glass knives
40
Q

resin blocks for UTSM and for EM; knife is expensive

A

diamond-edge

41
Q

for clearing and semi-thin sections

A

glass knives

42
Q

Used for modern lab, cheaper and convenient, composed of conventional steel knives

A

Disposable blades

43
Q

Microtome knives angle

A
  • clearance angle
  • wedge angle
  • bevel angle
  • rake angle
44
Q

Between the edge of the knife and the tissue block

A

Clearance angle

45
Q

Angle of cutting

A

Wedge angle

46
Q

Angle of cutting facet

A

Bevel angle

47
Q

(CA-BA) - 90°

A

Rake angle

48
Q

Angle of CA

A

5-15°

49
Q

Angle of WA

A

15°

50
Q

Angle of BA

A

27-32°

51
Q

Tests for microtome knife shapening

A
  • should cut a paraffin wax block at 2-4 um thickness w/o serrations when examined under the microscope (100x)
  • von mhol’s criterion
  • will split a hair drawn across it with only their own resistance
52
Q

Test that confirm shapening of knife is good when there is a Straight narrow, continuous life reflected from the knife edge until the other end if light is focused on the knife

A

Von Mhol’s criterion

53
Q

To remove nicks and irregularities

A

Honing

54
Q

Hones

A
  • Carborundum
  • arkansas
  • Belgium yellow
  • Belgian black vein
55
Q

Most expensive hone

A

Belgium yellow

56
Q

For rough side of honing

A
  • carborundum
  • arkansas
57
Q

For smooth side of honing

A

Belgium yellow

58
Q

T/f the finer the grain the harder the hone

A

T

59
Q

Used for final polishing

A

Diamantine

60
Q

After repeated sharpening, widened bevel angle (more than 35°)

A

Factory grinding

61
Q

T/f hones utilizes oilstones

A

T

62
Q

Material for honing

A

Smooth stones and machine hone

63
Q

To remove burrs and for final polishing

A

Stropping

64
Q

Lubricant for honing

A
  • soapy water, oil (mineral oil, castor oil, clove oil) or xylene
65
Q

Lubricant for stropping

A

Vegetable oil

66
Q

Direction for honing

A

Heel to toe

67
Q

Direction for stropping

A

Toe to heel

68
Q

Motion and no. Of strokes for honing

A

-zigzag
- 10-20 single strokes

69
Q

Motion and no. Of strokes for stropping

A

-zigzag
- 40-120 double strokes

70
Q

Flatten tissues in water bath at 10C lower than MP of paraffin wax

A

Floating

71
Q

What is used for adhesion

A

Mayer’s egg albumin

72
Q

Composition of Mayer’s egg albumin

A

-egg white
- glycerin
- thymol

73
Q

Transfer of tissue sections/ribbons on the slide using camel’s hair brush/applicator stick

A

Fishing out

74
Q

Correct positioning of the tissue section or ribbon on the slide

A

Orientation

75
Q

General steps in fixing sections onto slide

A
  1. Floating
  2. Adhesion
  3. Fishing out
  4. Orientation
  5. Deparaffinization
  6. Drying sections
  7. Post- mordantinh
76
Q

To remove excess paraffin

A

Deparaffinization

77
Q

Equipment for deparaffinization

A
  • alcohol lamp
  • paraffin oven
78
Q

Tissue will adhere to glass slide for long duration for staining

A

Drying sections

79
Q

Equipment for drying sections

A
  • wax oven
  • incubator
    -hot plate
  • alcohol lamp/ bunsen burner
80
Q

Temp for wax oven for drying sections

A

56-60 C for 2 hrs

81
Q

Temp for hot plate for drying sections

A

45-55C for 30-45 mins (fastest)

82
Q

To improve the demo of elements in the tissue in staining

A

Post- mordanting

83
Q

Secondary fixation(post-chroming) and used primarily as mordant and secondary fixative

A

Post - mordanting