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Histopath Lec > Sectioning > Flashcards

Sectioning Flashcards

1
Q

embedded tissue is trimmed and cut into uniformly thin slices using a microtome

A

Sectioning

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2
Q

The microtome consists of 3 essential parts, namely:

A

Block Holder (Chuck holder)

Knife carrier and knife

Pawl, Ratchet Feed Wheel and Adjustment screws-

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3
Q

where the tissue is held in position

A

Block Holder (Chuck holder)

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4
Q

for actual cutting of tissue sections

A

Knife carrier and knife

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5
Q

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

A

Pawl, Ratchet Feed Wheel and Adjustment screws

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6
Q

▫ Simplest Microtome
▫ For serial sections of large paraffin
embedded tissue

10-12 μ

A

Rocking Microtome (Cambridge Microtome)

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7
Q

▫ Most common type/ routinely used
▫ For routine paraffin embedded tissues
and Research laboratory

4-6 μ

A

Rotary Microtome

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8
Q

▫ Most dangerous type
▫ Recommended for celloidin, ester wax
blocks

7-9 μ

A

Sliding Microtome

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9
Q

2 types of Sliding Microtome:

A

a. Standard Sliding
b. Base Sledge/Sledge Type

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10
Q

▫ More dangerous
▫ Movable part is the knife for extremely
hard & rough tissues

7-9 μ

A

Standard Sliding

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11
Q

▫ Prefer to use
▫ Movable part is the block holder

7-9 μ

A

Base Sledge/Sledge Type

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12
Q

▫ Uses intermittent burst of CO2 that will freeze the block
▫ For cutting undehydrated tissues, neurological tissues, demonstration of fats

10-15 μ

A

Freezing Microtome

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13
Q

used as a propellant in Freezing Microtome

A

CO2

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14
Q

▫ For fresh tissue microtome
▫ For fluorescent antibody staining or histochemical enzyme studies

4μ

A

Cryostat or Cold Microtome

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15
Q

Cryostat or Cold Microtome
*tissues are refrigerated: ( __________)
*average _______

A

*tissues are refrigerated: ( -5 °C to –30 °C ) *average –20 °C

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16
Q

▫ For electron microscopy
▫ Uses diamond knife

0.5-1 μ

A

Ultrathin Microtome

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17
Q

Microtome Knives
-Mostly made up of

A

stainless steel

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18
Q

Plane Concave

SIDE

USE

LENGTH

A

Flat & Concave
*Less Concave- for celloidin *More concave- for paraffin

Base Sledge Rotary Microtome Rocking Microtome

25 mm

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19
Q

Biconcave

SIDE

USE

LENGTH

A

Both sides are Concave

Rotary Microtome

120 mm

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20
Q

Plane Wedge

SIDE

USE

LENGTH

A

Both sides are Flat

Base Sledge/
Sliding Microtome
*For Frozen section/ very hard tissues

100 mm

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21
Q

Bevel angle

A

27-32°

22
Q

Cutting angle

A

15°

23
Q

Clearance angle

A

0-15°

24
Q

Rake angle

A

Not stated

25
Q

Wedge angle

A

14-15°

26
Q

-KNIFE HARD SHARPENING

-grinding the cutting edges to acquire even size edge

Purpose: to remove blemishes & grinding the cutting

A

Honing

27
Q

Honing movement

A

HEEL to TOE MOVEMENT

28
Q

How many strokes in honing?

A

Uses 10-20, 20-30 strokes

29
Q

Hones/Oil stones used in honing (3)

A

a. Belgium Yellow: gives best results
b. Arkansas: with more polishing effects
c. Fine Carborundum: for badly nicked knives

30
Q

Lubricants used in honing (4)

A

a. Xylene/liquid paraffin
b. Mineral oil
c. Clove oil
d. Soapy water

31
Q

removal of the tissue ribbon from the float out bath

A

Fishing out

32
Q

Flotation Water Bath: temperature

A

5-10 °C lower than the melting point or 45-50 °C

33
Q

-POLISHING
Purpose: removal of burrs (irregularity of knife formed after honing)

A

Stropping

34
Q

Movement for stroping

A

TOE to HEEL MOVEMENT

35
Q

How many strokes for stroping?

A

40-120 double Strokes

36
Q

Stroping Uses PADDLE STROP made of horse leather that are usually treated with _________ oil or ________ oil at the back and not on the surface

A

Uses PADDLE STROP made of horse leather that are usually treated with Vegetable oil or Castor oil at the back and not on the surface

37
Q

Types of Sections:

A

Paraffin sections
Celloidin sections
Renal biopsy sections
Ultra-thin sections
Frozen section (cryostat)

38
Q

Sizes of different sections:

Paraffin sections
Celloidin sections
Renal biopsy sections
Ultra-thin sections
Frozen section (Cryostat)

A

o Paraffin sections: 4-6 μm sections (average: 5)

o Celloidin Sections: 10-15 μm sections

o Renal Biopsy sections: 2 μm section
*Semi-thin sections using plastic embedding medium

o Ultra-thin sections (EM)
*Recommended section: 80nm (silver or straw colored sections)

o Frozen section (Cryostat): 4 μm if microtome used is Rotary

39
Q

Drying Technique for Slides (3)

A
  1. Leave the slide in 37 °C incubator overnight
  2. Place the slide in oven for 2 hrs in 56-60°C
  3. Use a hot plate with 45-55°C for 30-45 minutes
40
Q

Adhesives (10)

A

Mayer’s Egg Albumin
Dried Albumin
1% Gelatin
Gelatin-Formaldehyde Mixture
Starch Paste
Plasma/Pooled Serum
Poly-L-Lysine
APES or 3-aminopropylthriethoxysilane
Sodium Silicate
Resins

41
Q

Most common
Equal amount of Egg white and Glycerin

A

Mayer’s Egg Albumin

42
Q

Dried albumin + sodium chloride
Addition of THYMOL CRYSTALS to prevent growth of molds

A

Dried Albumin

43
Q

Composed of Gelatin, Distilled water, Glycerol and phenol crystals Provides firmer attachment than albumin
Added to the flotation water bath
Disadvantage: Stains with many dyes

A

1% Gelatin

44
Q

1% Gelatin + 2% Formaldehyde

A

Gelatin-Formaldehyde Mixture

45
Q

Powdered starch, distilled water and HCL
Addition of thymol crystals to prevent growth of molds

A

Starch Paste

46
Q

Readily available from outdated blood stored in blood banks

A

Plasma/Pooled Serum

47
Q

Effective as an adhesive, slowly decreases in time Widely used as an adhesive in immunohistochem

A

Poly-L-Lysine

48
Q

Recommended for cytology

A

APES or 3-aminopropylthriethoxysilane

49
Q

Commercial syrup=1:10 dilution Had strong adhesive properties

A

Sodium Silicate

50
Q

Greatest adhesion

A

Resins

Histopath Lec (4 decks)

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