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MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE > LESSON 7: SECTIONING AND ADHESION OF TISSUE SECTIONS > Flashcards

LESSON 7: SECTIONING AND ADHESION OF TISSUE SECTIONS Flashcards

1
Q

✓ a previously processed tissue, is trimmed and cut into uniformly thin slices or sections

A

SECTIONING

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2
Q

✓ this procedure is also referred to as microtomy

A

SECTIONING

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3
Q

SECTIONING 3 BASIC PARTS

A
  1. BLOCK HOLDER
  2. KNIFE CARRIER AND KNIFE
  3. PAWL, RATCHET FEED WHEEL AND ADJUSTMENT SCREWS
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4
Q

TYPES OF MICROTOME

A

ROCKING

ROTARY

SLIDING

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5
Q

KINDS MICROTOME KNIVES

A

Light Microscopy
Plane-Concave, Plane-Wedge, Biconcave

Electron Microscopy
Diamond-edge , Broken Glass Knives

Others
Disposable blades , Safety razor blades

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6
Q

ANGLES DURING SECTIONING

A
  1. Clearance angle (5-10 degrees)
  2. Wedge angle (15 degrees)
  3. Bevel angle (27-32 degrees)
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7
Q

OTHER HONING PROCEDURES

A

❑ Plate Glass Honing

❑ Factory Grinding

❑ Automatic hones

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8
Q

GENERAL STEPS IN FIXING SECTIONS ONTO SLIDE

A
  1. Adhesion
  2. Floating on water bath
  3. Fishing out
  4. Orientation
  5. Deparaffinization and Drying Sections
  6. Post-mordanting
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9
Q

Adhesives:

A

 Mayer’s egg albumin (egg white + glycerin + thymol)

 Dried Albumin (dried albumin + NaCl + thymol)

 1% Gelatin (gealtin + glycerol + phenol)

 gelatin-formaldehyde mixture (gealtin + formaldehyde)

 poly-L-lysine

 APES (3-aminoprophylthriethoxysilane)

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10
Q

: where the embedded tissue is held in position

A
  1. BLOCK HOLDER
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11
Q

: perform the actual cutting of tissue sections

A
  1. KNIFE CARRIER AND KNIFE
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12
Q

: line up tissue block in proper position with the knife, adjust proper thickness

A
  1. PAWL, RATCHET FEED WHEEL AND ADJUSTMENT SCREWS
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13
Q

is brought in 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

spring-balanced teeth (pawl)

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14
Q

10-12 um

A

ROCKING

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15
Q

Paldwell Trefall

A

ROCKING

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16
Q

*simplest microtome

A

ROCKING

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17
Q

*for small and large paraffin-embedded blocks

A

ROCKING

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18
Q

*not for serial sections

A

ROCKING

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19
Q

4-6 um

A

ROTARY

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20
Q

Minot

A

ROTARY

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21
Q

*most common microtome

A

ROTARY

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22
Q

*for paraffin embedded blocks

A

ROTARY

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23
Q

4-9 um

A

SLIDING

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24
Q

Adams

A

SLIDING

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25
Q

*most dangerous

A

SLIDING

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26
Q

STANDARD SLIDING:

A

celloidin

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27
Q

BASE-SLEDGE:

A

hard and tough tissue blocks in all forms of media

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28
Q

25 mm

A

Plane-Concave

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29
Q

Plane side → celloidin
Concave → paraffin

A

Plane-Concave

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30
Q

100 mm

A

Plane-Wedge

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31
Q

Frozen sections

A

Plane-Wedge

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32
Q

Extremely hard and tough specimens in paraffin

A

Plane-Wedge

33
Q

120 mm

A

Biconcave

34
Q

Paraffin section

A

Biconcave

35
Q

Resin blocks

A

Diamond-edge

36
Q

Trimming

A

Broken Glass Knives

37
Q

Semi-thin sectioning

A

Broken Glass Knives

38
Q

Sharp cutting edge capable of producing 2-4 um sections

A

Disposable blades

39
Q

10 um and above

A

Safety razor blades

40
Q

Partially calcified tissues, paraffin and frozen sections

A

Safety razor blades

41
Q

✓ between the edge of the knife & the tissue block

A
  1. Clearance angle (5-10 degrees)
42
Q

✓ angle of cutting

A
  1. Wedge angle (15 degrees)
43
Q

✓ angle of cutting facet

A
  1. Bevel angle (27-32 degrees)
44
Q

Removal of gross nicks and blemishes

A

HONING

45
Q

Removal of burrs, for final polishing and sharpening of the cutting edge

A

STROPPING

46
Q

Smooth stones

A

HONING

47
Q

: best result

A

Belgium yellow

48
Q

: more polishing effect

A

Arkansas

49
Q

: coarsest

A

Fine carborundum

50
Q

Soapy water, Oil (Mineral oil, Castor oil, Clove oil) , Xylene, or liquid paraffin

A

HONING

51
Q

Vegetable Oil

A

STROPPING

52
Q

Heel-to-Toe, edge first

A

HONING

53
Q

Toe-to-Heel, edge last

A

STROPPING

54
Q

20-30 strokes in each direction

A

HONING

55
Q

10-20 strokes each surface (Minot and Plane-wedge knives)

A

HONING

56
Q

40-120 double strokes

A

STROPPING

57
Q

40-120 double strokes

A

STROPPING

58
Q

Horse leather

A

STROPPING

59
Q

➢ Diamantine: used for final polishing

A

❑ Plate Glass Honing

60
Q

➢ After repeated sharpening

A

❑ Factory Grinding

61
Q

➢ Widened Bevel angle (> 35O)

A

❑ Factory Grinding

62
Q

: performed to prevent washing out of tissue sections during staining

A

Adhesion

63
Q

is a substance which can be smeared onto the slides so that sections stick well to the slides

A

❖ Adhesive

64
Q

➢ Most commonly used (easy to prepare, convenient and relatively inexpensive)

A

 Mayer’s egg albumin (egg white + glycerin + thymol)

65
Q

➢ Sections are dried and stored in 70% alcohol until it is ready for staining

A

 Dried Albumin (dried albumin + NaCl + thymol)

66
Q

➢ Added to the waterbath during flotation rather than applying it on slides

A

 1% Gelatin (gealtin + glycerol + phenol)

67
Q

➢ slides are coated and allowed to dry at 37 OC for one hour or overnight prior to use

A

 gelatin-formaldehyde mixture (gealtin + formaldehyde)

68
Q

➢ aqueous detergent diluted to 0.01% concentration

A

 poly-L-lysine

69
Q

➢ widely used in immunohistochemistry

A

 poly-L-lysine

70
Q

➢ Very useful in cytology, particularly for cytospin preparations of proteinaceous or bloody material

A

 APES (3-aminoprophylthriethoxysilane)

71
Q

Floating on water bath: [?] LOWER than the MP of wax; to straighten and tissue sections

A

45-50 OC or 6-10 OC

72
Q

Fishing out: waterbath should have a temperature that is [?] lower than the melting point of the wax

A

10 OC

73
Q

: correct positioning of the tissue section/ribbon on the slide

A

Orientation

74
Q

Deparaffinization and Drying Sections
✓ wax oven (?)
✓ Incubators (?)
✓ Hot plate (?)
✓ Alcohol lamp/ bunsen flame
✓ Delicate tissues: [?]
✓ Blower-type electric slide dryer (?)

A

✓ wax oven (56OC – 60OC for 2 hrs)
✓ Incubators (overnight)
✓ Hot plate (45OC – 55OC for 30 – 45 mins.)
✓ Alcohol lamp/ bunsen flame
✓ Delicate tissues: 37 OC for at least 24 hours
✓ Blower-type electric slide dryer (50 – 55 OC for 20 – 30 mins.)

75
Q

✓ Secondary fixation (post-chroming)

A

Post-mordanting

76
Q

✓ 2.5 – 3% K2CrO4 for 24 hrs

A

Post-mordanting

77
Q

✓ Used primarily as mordant & secondary as fixative

A

Post-mordanting

78
Q

Post-mordanting
✓ 5 – 10 mins. in either

A

➢ Aq. Sol’n. of HgCl2
➢ Aq. Picric acid

79
Q

Post-mordanting
✓ 5 – 10 mins. in either

A

➢ Aq. Sol’n. of HgCl2
➢ Aq. Picric acid

MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE (9 decks)

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