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HISTOPATH MTAP > EMBEDDING - TRIMMING - SECTIONING - MICROTOME > Flashcards

EMBEDDING - TRIMMING - SECTIONING - MICROTOME Flashcards

(75 cards)

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1
Q

Aka: Casting/Blocking

A

EMBEDDING

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2
Q

● Process by which the impregnated tissue is placed into a precisely arranged position in a mold containing a medium which is then allowed to solidify

A

EMBEDDING

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3
Q

○ Process by which a tissue is arranged in precise positions in the mold during embedding,
■ On the microtome before cutting and on the slide before staining

A

● Orientation

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4
Q

The surface of the section to be cut should be placed parallel to the bottom of the mold in which it is oriented

A

EMBEDDING

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5
Q

Temperature of melted paraffin used for embedding

A

5-10℃ above its Melting Point

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5
Q

What is the other name of “Embedding”?

A

According to Gregorios it is otherwise
known as “Casting” or “Blocking”

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6
Q

To solidify embedded tissue

A

Cooled rapidly in a ref (-5℃) or immersed in cold water

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7
Q

STEPS IN EMBEDDING

A
  1. Removed the tissue from the cassette
  2. Fill the mold with wax
  3. Orientate the tissue
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8
Q

● Process of removing excess wax after embedding.

A

TRIMMING

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8
Q

is performed in the paraffin oven.

A

Embedding

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9
Q

● The excess wax is cut off from the block to expose the tissue surface in preparation for actual cutting.
● Knife/Blade may be used.

A

TRIMMING

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10
Q

Correct shape of the block:

A

Four-sided prism or Truncated pyramid

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11
Q

● Also known as cutting or microtomy

A

SECTIONING

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12
Q

● The process by which a processed tissue is cut into
uniformly thin slices (sections) to facilitate studies under the microscope.

○ Sections (Gregorio’s) or Ribbon

A

SECTIONING

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13
Q

Routine tissue microtomy

A

3 - 5 u

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14
Q

Frozen sections

A

10 - 15 u

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14
Q

Very thin sections
- For electron microscopy

A

60 - 100 nm

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15
Q

Semi-thin sections
- For electron microscopy

A

0.5 - 1 um

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15
Q

● For cutting serial sections of large blocks of paraffin-embedded tissues
● The simplest microtome

A

Rocking Microtome

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16
Q

Inventor of Rocking Microtome

A

Paldwell Trefall in 1881

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17
Q

Disadvantage:

● Difficulty in re-orienting the block

A

Rocking Microtome

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18
Q

● For cutting paraffin-embedded
sections

A

Rotary / Minot Microtome

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19
Q

Inventor of Rotary / Minot Microtome

A

Minot in 1885-1886

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20
Q

● The most common microtome used today especially for paraffin-embedded tissues

A

Rotary / Minot Microtome

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21
Some microtomes have digital indicators to adjust the thinness of the sections to be cut.
Rotary / Minot Microtome
22
● For cutting celloidin-embedded sections
Sliding Microtome
23
Inventor of Sliding Microtome
Adams in 1789
24
The most dangerous type due to movable exposed knife
Sliding Microtome
25
2 types of Sliding Microtome:
1. Base-Sledge 2. Standard Sliding Microtome
26
can be used for all types of media - Block holder: moving - Knife: stationary
Base-Sledge
27
- Block holder: stationary - Knife: moving
Standard Sliding Microtome
28
It is another type of microtome
Rotary Rocking Microtome
29
Disadvantage: ● Sections are liable to disintegrate
Vibratome
29
● Used for unfixed, unfrozen specimen sectioning for enzyme demonstrations
Vibratome
30
● For cutting sections for electron microscopy
Ultrathin Microtome
31
Temperature: Ultrathin Microtome
0-4 degree celsius
32
Impregnating medium of Ultrathin Microtome
Plastic or resin impregnating media
33
Uses diamond knives or broken plate glass ○ Broken plate glass is only used as an ALTERNATIVE to diamond knives
Ultrathin Microtome
34
● For cutting of unembedded frozen sections
Freezing Microtome
34
Plane Concave Usual Length: Description:
25 mm One side ● flat Other side ● concave
34
● Specimen is small, fixed in osmium tetroxide, embedded in plastic
Ultrathin Microtome
35
Among the two types of sliding microtome, which is the most dangerous type?
Standard sliding microtome is the most dangerous type because of the exposed movable knife
35
Inventor of Freezing Microtome
Queckett in 1848
36
Plane Wedge Usual Length: Description:
100 mm Both sides ● straight
36
Biconcave Usual Length: Description:
120 mm Both sides ● concave
37
● How far the knife is from the tissue block
Clearance Angle
38
Clearance Angle
0 to 15 degrees
39
Bevel Angle
27 to 32 degrees
40
● Types of Hones (3)
○ Belgium Yellow ○ Fine Carborundum ○ Arkansas
40
● Removal of gross nicks
Honing
41
gives the best honing result
Belgium Yellow
42
● Knife direction: ○ Heel to toe
Honing
43
● Removal of burr
Stropping
44
● Knife direction: ○ Toe to heel
Stropping
45
● Instrument: ○ Paddle strop
Stropping
46
made up of horse leather
Paddle strop
47
TEMPERATURE OF FLOATATION WATER BATH (2)
● 45-50 degrees Celsius ● 6-10 degrees Celsius LOWER than the melting point of wax
47
Why LOWER than the melting point of wax
If the temperature is higher than the melting point, the ribbons will melt, and the tissues will submerge to the bottom
48
Electron Microscopy
0-10°C
49
Fixation Surgical Specimen
25°C
50
Paraffin Oven for Impregnation
55-60°C or 2-5°C ABOVE the melting point of paraffin wax
51
Embedding
5-10°C ABOVE the melting point of paraffin wax
52
Microwave Fixation
45-55°C
53
Most common adhesive agent
Mayer’s Egg Albumen
54
Components of Mayer’s Egg Albumen (2)
○ Egg white/Albumen ○ Crystals of Thymol
55
to prevent growth of molds
Crystals of Thymol
56
Used to enhance tissue adherence to slides
Dried Albumin
57
Commonly used for coating slides to improve tissue section adhesion
Gelatin
58
Provides strong adhesion and prevents tissue loss during staining
Gelatin-Form aldehyde Mixture
59
Simple and effective adhesive for mounting specimens
Starch paste
60
Effective natural adhesive agent for tissue sections
Plasma
61
Enhances cell attachment, particularly useful for immunohistochemistry
Poly-L-Lysine
62
3-APES meaning
3-aminopropyltriethoxysilane
63
● APES-coated slides ○ very useful in cytology, particularly for cytospin preparations of proteinaceous or bloody material
3-APES (3-aminopropyltriethoxysilane)
64
(2) STAGES OF KNIFE SHARPENING
(1) Honing (2) Stropping

HISTOPATH MTAP (5 decks)

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