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basic histologic techniques Flashcards

1
Q

12 Steps of Histologic Techniques

A 
numbering
receiving of specimens
fixation
dehydration
clearing
infiltration
embedding
blocking and trimming
sectioning
staining
mounting
labelling
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2
Q

first and most important step

logging in a log book the details of the tissue

A

numbering

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3
Q

tissue specimens should be properly labeled and with a corresponding request
tissue s will be described by the pathologist (gross description)
tissues will be cut by the pathologist

A

receiving of specimens

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4
Q

tissues are immersed in a fixative to prevent decomposition and preserve structure

A

fixation

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5
Q

removal of excess water using ascending grades of alcohol

A

dehydration

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6
Q

removal of excess alcohol
makes tissues clear/transparent
prepares tissues for paraffin impregnation

A

clearing/dealcoholization

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7
Q

tissue cavities are saturated w/ paraffin wax

A

infiltration

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8
Q

impregnated tissue is oriented and embedded in paraffin block to form a tissue block

A

embedding

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9
Q

separation of embedded tissues into blocks

removal of excess paraffin wax using knives

A

blocking and trimming

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10
Q

very thin sections of tissue are cut using microtome

A

sectioning

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11
Q

using H& E (routine stain)

A

staining

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12
Q

stained tissue sections on a slide are added w/ mounting medium and covered w/ coverslip

A

mounting

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13
Q

final step in tissue preparation

A

labeling

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13
Q

Responsible for transcribing the dictations of the pathologist

A

Medical technologist

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14
Q

Used in labeling slecimens

A

Pencil

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15
Q

Provides the gross description

Cuts of sections the specimen when received

A

Pathologist

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16
Q

Technique done to kill microorganism in the specimen

A

Fixation

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17
Q

Routinely used fixative

A

Formalin (10%) or formaldehyde

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18
Q

Stock solution of formalin that must be diluted

A

37% formalin

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19
Q

Agent used in dehydration

A

Increasing grades of Ethanol or ethyl alcohol

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20
Q

Effect of clearing to tissues

A

Tissues become translucent or transparent

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21
Q

Most common clearing agent

A

Xylol or xylene

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22
Q

Melting point of paraffin wax

A

56-57C

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23
Q

Ways of performing infiltration

A

Manual

Using automatic tissue processor

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24
Another name for infiltration
Impregnation
25
Performs fixation to infiltration (steps 2-5) | Has a basket wherein tissue cassette will be placed
Automatic tissue processor
26
How long do cutting to infiltration take?
1 day
27
The act of placing impregnated tissue in the embedding mold
Orientation
28
Examples of embedding mold
Paper boat Plastic mold Ice cube tray
29
Separation of embedded tissues into blocks | Separating small block from big block
Blocking
30
Removal of excess paraffin wax using knives until a truncated pyramid is made
Trimming
31
Thickness required when sectioninf
4-6 nm
32
Where tissue section is placed before in the microtome Surface is colored black Removes wrinkles
Flotation Water bath
33
After flotation, perform ____
Fishing
34
Act of scooping the tissue from the flotation water bath
Fishing out
35
Another term for staining
Coloring
36
A nuclear stain | Color is blue or dark blue
Hematoxylin
37
A cytoplasmic stain | Reddish or pinkish
Eosin
38
Also serves as a counterstajn
Eosin
39
Provides contrast and background
Eosin
40
2 types of mounting medium
Aqueous | Resinous
41
Primary mounting medium used in histology
Resinous
42
Ex of aqueous mounting medium
Water
43
Example of resinous mounting medium
Eukitt and Canada balsam
45
Final step Use of gum labels May be computerized or bar coded
Labelling
46
– fixative often used for EM; reinforces fixation by being a dialdehyde capable also of cross-linking proteins
• Gluteraldehyde
47
– preserves and stains membrane lipids and proteins
Osmium tetroxide
48
In electron microscopy, a _____ procedure is done.
double-fixation
49
basic histologic technique • ethanol is then replaced by an organic solvent miscible with both alcohol and embedding medium • Alcohol is removed in toluene or other agents in which both alcohol and paraffin are miscible
3. Clearing
50
* Fully cleared tissue is placed in melted paraffin in an oven at 52 – 60C * At 52 – 60C, clearing solvent evaporates and tissue is filled with paraffin wax
4. Infiltration
51
* The paraffin-infiltrated tissue is placed in a small mold with melted paraffin and allowed to harden * Impregnated tissue hardens in a small container of paraffin at RT
Embedding
52
SPATIAL UNITS in Histology:
micrometer, nanometer, angstrom
53
– cell components with a net negative charge (anionic) stain readily with basic dyes (eg. Nucleic acids) -have acids in their composition
• Basophilic
54
- cell components that are cationic have affinity for acidic dyes (eg. Proteins with many ionized amino groups)
• Acidophilic
55
– toluidine blue, alcian blue, methylene blue, hematoxylin
• Basic Dyes
56
– eosin, orange G, acid fuchsin
• Acid dyes
57
type of stain – used in more complex histologic procedures -help to distinguish extracellular tissue components better than H&E
• Trichrome (Mallory stain, Masson stain)
58
chromatin with active dna | stains lightly
euchromatin
59
inactive dna | stains darkly
heterochromatin
60
euchromatin that becomes a heterochromatin
facultative heterochromatin
61
whole process of an X chromosome becoming a barr body or sex chromatin
lyonization
62
according to lyon's hypothesis, at the _____ cell stage, one of the X in the female gets turned off
100
63
specimen used for determining sex chromatin
``` peripheral blood (neutrophil) tongue and buccal cavity ```
64
first blood must be wiped off as it contains
tissue juices
65
methanol number of seconds
5 seconds, air dry
66
eosin number of seconds
3
67
methylene blue number of seconds
6 seconds

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