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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
*most dangerous
SLIDING
26
STANDARD SLIDING:
celloidin
27
BASE-SLEDGE:
hard and tough tissue blocks in all forms of media
28
25 mm
Plane-Concave
29
Plane side → celloidin Concave → paraffin
Plane-Concave
30
100 mm
Plane-Wedge
31
Frozen sections
Plane-Wedge
32
Extremely hard and tough specimens in paraffin
Plane-Wedge
33
120 mm
Biconcave
34
Paraffin section
Biconcave
35
Resin blocks
Diamond-edge
36
Trimming
Broken Glass Knives
37
Semi-thin sectioning
Broken Glass Knives
38
Sharp cutting edge capable of producing 2-4 um sections
Disposable blades
39
10 um and above
Safety razor blades
40
Partially calcified tissues, paraffin and frozen sections
Safety razor blades
41
✓ between the edge of the knife & the tissue block
1. Clearance angle (5-10 degrees)
42
✓ angle of cutting
2. Wedge angle (15 degrees)
43
✓ angle of cutting facet
3. Bevel angle (27-32 degrees)
44
Removal of gross nicks and blemishes
HONING
45
Removal of burrs, for final polishing and sharpening of the cutting edge
STROPPING
46
Smooth stones
HONING
47
: best result
Belgium yellow
48
: more polishing effect
Arkansas
49
: coarsest
Fine carborundum
50
Soapy water, Oil (Mineral oil, Castor oil, Clove oil) , Xylene, or liquid paraffin
HONING
51
Vegetable Oil
STROPPING
52
Heel-to-Toe, edge first
HONING
53
Toe-to-Heel, edge last
STROPPING
54
20-30 strokes in each direction
HONING
55
10-20 strokes each surface (Minot and Plane-wedge knives)
HONING
56
40-120 double strokes
STROPPING
57
40-120 double strokes
STROPPING
58
Horse leather
STROPPING
59
➢ Diamantine: used for final polishing
❑ Plate Glass Honing
60
➢ After repeated sharpening
❑ Factory Grinding
61
➢ Widened Bevel angle (> 35O)
❑ Factory Grinding
62
: performed to prevent washing out of tissue sections during staining
Adhesion
63
is a substance which can be smeared onto the slides so that sections stick well to the slides
❖ Adhesive
64
➢ Most commonly used (easy to prepare, convenient and relatively inexpensive)
 Mayer’s egg albumin (egg white + glycerin + thymol)
65
➢ Sections are dried and stored in 70% alcohol until it is ready for staining
 Dried Albumin (dried albumin + NaCl + thymol)
66
➢ Added to the waterbath during flotation rather than applying it on slides
 1% Gelatin (gealtin + glycerol + phenol)
67
➢ slides are coated and allowed to dry at 37 OC for one hour or overnight prior to use
 gelatin-formaldehyde mixture (gealtin + formaldehyde)
68
➢ aqueous detergent diluted to 0.01% concentration
 poly-L-lysine
69
➢ widely used in immunohistochemistry
 poly-L-lysine
70
➢ Very useful in cytology, particularly for cytospin preparations of proteinaceous or bloody material
 APES (3-aminoprophylthriethoxysilane)
71
Floating on water bath: [?] LOWER than the MP of wax; to straighten and tissue sections
45-50 OC or 6-10 OC
72
Fishing out: waterbath should have a temperature that is [?] lower than the melting point of the wax
10 OC
73
: correct positioning of the tissue section/ribbon on the slide
Orientation
74
Deparaffinization and Drying Sections ✓ wax oven (?) ✓ Incubators (?) ✓ Hot plate (?) ✓ Alcohol lamp/ bunsen flame ✓ Delicate tissues: [?] ✓ Blower-type electric slide dryer (?)
✓ 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
✓ Secondary fixation (post-chroming)
Post-mordanting
76
✓ 2.5 – 3% K2CrO4 for 24 hrs
Post-mordanting
77
✓ Used primarily as mordant & secondary as fixative
Post-mordanting
78
Post-mordanting ✓ 5 – 10 mins. in either
➢ Aq. Sol’n. of HgCl2 ➢ Aq. Picric acid
79
Post-mordanting ✓ 5 – 10 mins. in either
➢ Aq. Sol’n. of HgCl2 ➢ Aq. Picric acid

MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE (9 decks)

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