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HISTOPATH > Lesson 1: CYTO AND HISTOTECHNIQUES (ADVANCE CLASS) > Flashcards

Lesson 1: CYTO AND HISTOTECHNIQUES (ADVANCE CLASS) Flashcards

1
Q

rooms in histopathology lab should have?

A

proper ventilation and good lighting

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2
Q

Wax is heated at ___ in the embedding
machine

A

60C

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3
Q

color of tissue cassette?
Skin

A

Gray

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4
Q

color of tissue cassette?
Lymph nodes

A

pink

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5
Q

Processes
that an automated machine can perform

A

FDCI (Fixation, Dehydration, Clearing, Infiltration)

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6
Q

sections produced by a microtome

A

Ribbons

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7
Q

color of tissue cassette?
-Liver/renal

A

Yellow

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8
Q

color of tissue cassette?
Routine (e.g. placenta/uterus)

A

Green

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9
Q

color of tissue cassette?
Bones

A

White

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10
Q

Fresh tissue examination procedure

A

A. Teasing or Dissociation
B. Squash Preparation
C. Smear Preparation

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11
Q

Smear Preparation procedure

A

➢ Streaking (zigzag manner)
➢ Spreading (spreading in a circular motion)
➢ Pull-apart
➢ Touch Preparation

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12
Q

FIXATION
Tissue-to-Fixative Ratio–

A

1:10 or 1:20

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13
Q

Accomplished by preserving and carefully processing
solid structures and tissues in the following order:

A
  1. Fixation
  2. Decalcification (OPTIONAL)
  3. Dehydration
  4. Clearing
  5. Impregnation (INFILTRATION)
  6. Embedding
  7. Trimming
  8. Microtomy
  9. Staining
  10. Mounting
  11. Labeling
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14
Q

Gas produced by oxidation of methanol

A

Formaldehyde

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15
Q

Folmaldehyde 100% concentration?

A

gas form

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16
Q

Folmaldehyde 37-40% concentration?

A

stock concentration (causes overhardening of the external surfaces of tissues)

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17
Q

working solution; MOST COMMONLY USED IN PH
-Takes longer to prepare

A

10% Neutral Buffered Formalin

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18
Q

10% Neutral Buffered Formalin
Fixation Time:

A

4-24 hours

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19
Q

BEST GENERAL TISSUE FIXATIVE

-Best for iron-containing pigments and elastic fibers
which do not stain well after Susa, Zenker, or
Chromate fixation

A

10% Neutral Buffered Formalin

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20
Q

Thickness of Specimen

○ Light Microscope:___
○ Electron Microscope: _____

A

2 cm2 x 0.4 cm2
1-2 mm2

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21
Q

Temperature
○ Room Temperature ______: ROUTINE

A

(22C–28C)–40C

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22
Q

factors affecting fixation

A

● Fixative of choice
● Time (20-30 minutes)
● Tissue-to-Fixative Ratio
● Penetration Rate (1 mL/hr)
● Thickness of Specimen
● Tissue Components
● pH
● Temperature
● Osmolality
● Agitation, Vacuum

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23
Q

Temperature
○ Tissue Processors: ___

A

45C

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24
Q

Temperature
○ Microwave: ___

A

65C

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25
Q

Temperature
○ Tuberculosed Tissues: ____

A

100C

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26
Q

Temperature
○ Electron Microscope: ____

A

0C–40C

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27
Q

MOST COMMONLY USED ALCOHOL IN DEHYDRATION?

A

Ethanol

28
Q

Osmolality
○ MOST OPTIMAL: ______

A

400–450 mOsm
(milliosmole)

29
Q

Tissue-to-Dehydrating Agent Ratio

A

1:10

30
Q

Temperature
○ Rapid Biopsy: ___

A

60C

31
Q

● Removal of fixative and water from the tissue
● Carried out by passing the tissues through a series of
ascending grades of alcohol_________

A

Dehydration
70%, 90%, and
absolute alcohol

32
Q

Decalcification done for how many hours?

A

48 hours

33
Q

Removal of calcium ions from a bone or calcified tissue through a histological process that makes them flexible and easier to cut

A

Decalcification

34
Q

most commonly employed medium in
histopathology

A

Paraffin Wax

35
Q

Can cause damage to
knife, poor trimming of tissues

A

INADEQUATELY DONE

36
Q

Tissue-to-Decalcifying Agent Ratio

A

1:120

37
Q

factors influencing the rate of decalcification

A
  1. CONCENTRATION
  2. FLUID ACCESS
  3. SIZE AND CONSISTENCY
  4. AGITATION
  5. TEMPERATURE
38
Q

Completely transparent tissue indicates the
_____ of the clearing process

A

TERMINAL POINT

39
Q

● AKA dealcoholization
● Process of removing the dehydrating agent from the tissue to facilitate impregnation of paraffin wax
● MAIN GOAL: Make tissues transparent/translucent

A

clearing

40
Q

FROZEN SECTIONS can use _____ and ____ as dehydrating agent

A

glycerin, gum
syrup

41
Q

xylene 1 and 2 time interval?

A

20 minutes

42
Q

xylene 3 time interval

A

45 minutes

43
Q

○ Commonly employed clearing agent; cheapest
○ Clear fluid as well as a cheap and excellent
lipid solvent
○ Becomes milky if tissue is not properly
dehydrated

A

Xylene

44
Q

Infiltration and embedding
Duration per Section:

A

1 hour–1 hour–3 hours

45
Q

-Impregnate in a medium that could support the tissue

-For easier handling of tissue

A

infiltration and embedding

46
Q

Infiltration and embedding
○ Wax/Paraffin Oven: ____ than melting
point
○ Wax can only be reused _____

A

> 2C–5C
twice

47
Q

INFILTRATION Procedure

Immerse the tissue samples in 3 stages of paraffin wax following the given time intervals respectively:
● 1st change paraffin wax bath: _____
● 2nd change paraffin wax bath: ______
● 3rd change paraffin wax bath: _____

A

30 minutes
30 minutes
45 minutes

48
Q

Paraffin wax may be used only twice
- Removed by heating the wax to _____

A

100-105C

49
Q

Temperature of embedding paraffin wax?

A

2-4C above its melting point

50
Q

● Arrangement of the tissue in a precise position in
the mold during embedding
● MOST CRUCIAL STEP DURING EMBEDDING

A

Orientation

51
Q

OVERHEATING OF WAX cause

A

○ Hardening and distortion of the tissue
○ Breakdown of the paraffinic additive (e..g
resin polymer)

52
Q

usually form ribbons due to slight heat
generated between the block and the knife edge during the process of cutting

A

SECTIONS

53
Q

Usual thickness of tissue in sectioning

A

10 microns

54
Q

AFTER MICROTOMY
Heat the slide gently on the slide warmer (_____) until the ribbons spread and flatten ou

A

40-45C

55
Q

Floatation Bath:____ lower than temperature of
paraffin wax

A

5-10C

56
Q

Spring-balance or pawl is brought into contact with the block, 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

Microtome

57
Q

used to clean the
knife

A

Xylene/Wax Paper/Brush

58
Q

Process of applying des on the section to see and study the architectural pattern of the tissue and physical characteristics of the cells

A

staining

59
Q

basic, attracted to acidic part (nucleus) of the cell

-Stains purple-blue

A

Hematoxylin Stain

60
Q

acidic, attracted to basic part
(cytoplasm) of the cell

-Stains red-orange

A

Eosin Stain

61
Q

Rehydration: descending grades of alcohol________

A

(100%–90%–70%)

62
Q

Routine H&E Staining procedure

A

STEP 1 Deparaffinization
STEP 2 Rehydration: Graded alcohol
STEP 3 Nuclear stain: Haematoxylin
STEP 4 Differentiation
STEP 5 Bluing
STEP 6 Counterstain: Eosin
STEP 7 Dehydration
STEP 8 Clearing and Mounting

63
Q

In Routine H&E Staining procedure what is the nuclear stain?

A

Haematoxylin

64
Q

In Routine H&E Staining procedure what is the counterstain?

A

eosin

65
Q

Process of removing excess hydrogen ion
from the stain

A

BLUING

66
Q

Mounting Protective layer:

A

between 1.0 and 2.5 mm thick

67
Q

Process of placing the coverslip on the slide with the use of a mounting media

A

Mounting

HISTOPATH (4 decks)

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