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Botany > basic histologic techniques > Flashcards

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
Q

Another name for infiltration

A

Impregnation

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25
Q

Performs fixation to infiltration (steps 2-5)

Has a basket wherein tissue cassette will be placed

A

Automatic tissue processor

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26
Q

How long do cutting to infiltration take?

A

1 day

27
Q

The act of placing impregnated tissue in the embedding mold

A

Orientation

28
Q

Examples of embedding mold

A

Paper boat
Plastic mold
Ice cube tray

29
Q

Separation of embedded tissues into blocks

Separating small block from big block

A

Blocking

30
Q

Removal of excess paraffin wax using knives until a truncated pyramid is made

A

Trimming

31
Q

Thickness required when sectioninf

A

4-6 nm

32
Q

Where tissue section is placed before in the microtome
Surface is colored black
Removes wrinkles

A

Flotation Water bath

33
Q

After flotation, perform ____

A

Fishing

34
Q

Act of scooping the tissue from the flotation water bath

A

Fishing out

35
Q

Another term for staining

A

Coloring

36
Q

A nuclear stain

Color is blue or dark blue

A

Hematoxylin

37
Q

A cytoplasmic stain

Reddish or pinkish

A

Eosin

38
Q

Also serves as a counterstajn

A

Eosin

39
Q

Provides contrast and background

A

Eosin

40
Q

2 types of mounting medium

A

Aqueous

Resinous

41
Q

Primary mounting medium used in histology

A

Resinous

42
Q

Ex of aqueous mounting medium

A

Water

43
Q

Example of resinous mounting medium

A

Eukitt and Canada balsam

45
Q

Final step
Use of gum labels
May be computerized or bar coded

A

Labelling

46
Q

– fixative often used for EM; reinforces fixation by being a dialdehyde capable also of cross-linking proteins

A

• Gluteraldehyde

47
Q

– preserves and stains membrane lipids and proteins

A

Osmium tetroxide

48
Q

In electron microscopy, a _____ procedure is done.

A

double-fixation

49
Q

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

A
  1. Clearing
50
Q
  • 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
A
  1. Infiltration
51
Q
  • 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
A

Embedding

52
Q

SPATIAL UNITS in Histology:

A

micrometer, nanometer, angstrom

53
Q

– cell components with a net negative charge (anionic) stain readily with basic dyes (eg. Nucleic acids)
-have acids in their composition

A

• Basophilic

54
Q
  • cell components that are cationic have affinity for acidic dyes (eg. Proteins with many ionized amino groups)
A

• Acidophilic

55
Q

– toluidine blue, alcian blue, methylene blue, hematoxylin

A

• Basic Dyes

56
Q

– eosin, orange G, acid fuchsin

A

• Acid dyes

57
Q

type of stain
– used in more complex histologic procedures
-help to distinguish extracellular tissue components better than H&E

A

• Trichrome (Mallory stain, Masson stain)

58
Q

chromatin with active dna

stains lightly

A

euchromatin

59
Q

inactive dna

stains darkly

A

heterochromatin

60
Q

euchromatin that becomes a heterochromatin

A

facultative heterochromatin

61
Q

whole process of an X chromosome becoming a barr body or sex chromatin

A

lyonization

62
Q

according to lyon’s hypothesis, at the _____ cell stage, one of the X in the female gets turned off

A

100

63
Q

specimen used for determining sex chromatin

A 
peripheral blood (neutrophil)
tongue and buccal cavity
64
Q

first blood must be wiped off as it contains

A

tissue juices

65
Q

methanol number of seconds

A

5 seconds, air dry

66
Q

eosin number of seconds

A

3

67
Q

methylene blue number of seconds

A

6 seconds

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