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Histopath LEC > IMPREGNATION > Flashcards

IMPREGNATION Flashcards

1
Q

Impregnation aka

A

Infiltration

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2
Q

IMPREGNATION (a.k.a. Infiltration)

• Process that removes the_____
• Fills the_____
• Permeates tissue with a____
• Incomplete impregnation -______

A

clearing agent

tissue cavities

support medium

tissue airholes

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3
Q

3 TYPES OF GENERAL TISSUE IMPREGNATION

A

Paraffin wax impregnation
Celloidin impregnation
Gelatin impregnation

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4
Q

Requires_____ changes of melted paraffin wax

A

2 or more

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5
Q

Kidney specimen

Ika ilan ang impregnation???

A

4th

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6
Q

Requires 2 different reagent for embedding

A

Double embedding

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7
Q

How many changes in infiltration if microwave oven is utilized?

A

4 changes (shorter time)

Ex. 15 or 20mins each

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8
Q

Which provides structure for sectioning? Infiltration or embedding

A

Infiltration

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9
Q

If may mali sa dehydration asa makita?

A

Clearing

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10
Q

If may mali sa infiltration asa makita?

A

Sectioning

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11
Q

If may mali sa clearing asa makita?

A

Sectioning

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12
Q

Paraffin wax

Melting point: 45°C, 52°C, 56°C, 58°C @RT (20-24°C)

Paraffin oven must be maintained_____ the melting point of wax

A

2-5°C above

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13
Q

If the MP if paraffin wax is 59, what is the max temperature of oven?

A

64

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14
Q

C to F formula

A

°F = (9/5 × °C) + 32

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15
Q

Paraffin wax

Used pure

Wax must be filtered first using coarse filter paper such as Green’s No. 904 in oven at 2°C higher than the melting point of wax.

Reusable only once, but heat it first @
________ (remove water)

A

100-105°C

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16
Q

PARAFFIN WAX

Advantages

A

• Rapid (24 hours)

• Maybe cut with ease without undue distortion

• Many staining procedures are permitted

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17
Q

PARAFFIN WAX

Disadvantages

A

• Prolonged impregnation will cause excessive shrinkage

• Not recommended to fatty tissues (ex. breast)

• Overheated paraffin makes the specimen brittle

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18
Q

3 Ways of Paraffin Wax Impregnation:

A

Manual processing
Automatic processing
Vacuum embedding

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19
Q

manual processing

This is what we are performing in the laboratory.

In this illustration, there are ______of paraffin; 1 hour per beaker/change.

The changes will depend on the protocol of the laboratory.

A

3 changes

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20
Q

automatic processing (MACHINE USED?)

• Our automatic processor is usually designed with 12 stations. The infiltration will take place up at stations 11 & 12.

The infiltration time will be faster because there is constant agitation in the machine.

Hence, a shorter processing time.

A

Autotechnicon & Elliott bench-type

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21
Q

Asa na step ma dissolve ang fats?

A

Clearing

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22
Q

3 Ways of Paraffin Wax Impregnation:

Where is agitation observed?

A

Manual and Automated processing

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23
Q

• Among the 3 processes, this type is the fastest. However, it is expensive because we need to acquire first the machine itself

A

vacuum embedding

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24
Q

vacuum embedding

it can only perform the________.
Unlike with our automatic processing, from________.

A

embedding/ infiltration process

Fixation down to infiltration.

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25
Q

Minimum capacity of canister in automated tissue processor

A

200 cassettes

Max: >200

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26
Q

How to know if reagent is still good: xylene

A

Not white

If white, may water

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27
Q

How to know if reagent is still good: alcohol in dehydration

A

Ignite

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28
Q

How to know if reagent is still good: infiltration ing media

A

Amoy wax pa

If amoy xylene, wala na

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29
Q

SUBSTITUTE FOR PARAFFIN WAX

A

PEBEW

Paraplast

Embeddol

Bioloid

Ester wax

Water Soluble Waxes (Polyethylene glycol)

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30
Q

Which substitute for paraffin wax have the same Mp

A

Paraplast
Embeddol

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31
Q

Mp of paraplast and embeddol

A

56-57

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32
Q

Mac temp for paraplast and embeddol

A

62

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33
Q

• semisynthetic; for embedding of eyes

A

Bioloid

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34
Q

Ester Wax
o Melting Point:

A

46-48°C

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35
Q

Mixture of pure paraffin and synthetic plastic polymer (Dimethyl sulfoxide); more elastic and resilient

A

Paraplast

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36
Q

Less brittle, and less compressible

A

Embeddol

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37
Q

Harder than paraffin thus used with sliding/sledge-type microtome

A

Ester wax

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38
Q

What type of microtome is used for ester wax?

A

Sliding microtome

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39
Q

Ester wax

o Water insoluble but soluble in______, thus prior______ is not needed

o But Cellosolve, and xylene may be used if indicated

A

95% ethanol; clearing

40
Q

Substitute for paraffin wax

  • most common
    • No need for dehydration and clearing thus sections are difficult to float out during the fishing method and mount
41
Q

Carbowax - most common
• No need for _________ thus sections are difficult to float out during the fishing method and mount

A

dehydration and clearing

42
Q

What reagent is used for fatty specimens? For viewing adipose tissues?

A

Carbowax (water soluble waxes)

43
Q

Water Soluble Waxes (Polyethylene glycol)

• Melting Point:

A

38-42°C or 45-56°C

44
Q

Carbowax

difficult to float out during the fishing method and mount

• Remedy:

A

add soap to water or

10% PEG 900 in water

45
Q

CELLOIDIN IMPREGNATION
• A.k.a

46
Q

CELLOIDIN IMPREGNATION
• A.k.a Colloidin

Used for

A

Large, hollow cavities

Bones
Brain
Teeth

47
Q

• Specimen with large and hollow cavities which tend to collapse; hard and dense tissues; neurologic tissues

A

Celloidin impregnation

48
Q

What type of microtome is used for large and hollow cavities

A

Sliding microtome

49
Q

• Purified form of nitrocellulose/gun cotton

50
Q

Celloidin

Concentration: in_________ dissolved in equal parts of ether and alcohol - increasing

A

2%, 4%, 8%

51
Q

CELLOIDIN
ADVANTAGES

A

• Does not require heat; less shrinkage
• Cutting of thicker tissues
• Recommended for neurological tissues

52
Q

CELLOIDIN

DISADVANTAGES

A

Very slow (days or week)
Thin sections are difficult to cut
Very volatile

53
Q

METHODS OF CELLOIDIN INFILTRATION (WET)

A

Fixation & Dehydration

Place tissue in ether-alcohol

Thin celloidin (2%)

Medium celloidin (4%)

Thick celloidin (8%)

Remove specimen and put in a fresh thick celloidin

Keep in jar or desiccator until ether-alcohol evaporates

WET: Store tissue block in 70%-80% alcohol

54
Q

METHODS OF CELLOIDIN INFILTRATION (DRY)

A

Fixation & Dehydration

Place tissue in ether-alcohol

Thin celloidin (2%)

Medium celloidin (4%)

Thick celloidin (8%)

Remove specimen and put in a fresh thick celloidin

Keep in jar or desiccator until ether-alcohol evaporates

DRY: Gilson’s Mixture (chloroform and cedarwood oil)

55
Q

Gilson’s Mixture

A

chloroform and cedarwood oil

56
Q
  • for bones, brain, teeth
    • Store tissue block in 70%-80% alcohol
    The purpose of storing it in this concentration is to avoid dehydration and shrinkage of the tissue.
A

Wet celloidin infiltration

57
Q

for whole eye sections
Store tissue block in Gilson’s Mixture (chloroform and cedarwood oil)

A

Dry celloidin infiltration

58
Q

• Has lower viscosity, thus can be used in higher concentration, and rapid tissue penetration

A

Nitrocellulose/Low Viscosity Nitrocellulose

59
Q

• Advantages: Harder tissues blocks, thus thinner sections are possible

• Disadvantages: Explosive when dry due to nitrates

A

Nitrocellulose/Low Viscosity Nitrocellulose

60
Q

It consists Plasticizer

• E.g. oleum ricini or castor oil
• Needed to prevent tissue cracking in chrome mordanted tissues
• This promotes plasticity as well as flexibilitv in order to reduce brittleness.

A

Nitrocellulose/Low Viscosity Nitrocellulose

61
Q

Nitrocellulose/Low Viscosity Nitrocellulose Luh

Advantages:
Disadvantages:

A

• Advantages: Harder tissues blocks, thus thinner sections are possible

• Disadvantages: Explosive when dry due to nitrates

62
Q

How to prevent nitrocellulose from exploding?

A

suspended with alcohol to maintain its liquid state

63
Q

Nitrocellulose

• E.g. oleum ricini or castor oil
• Needed to prevent tissue cracking in chrome mordanted tissues
• This promotes plasticity as well as flexibility in order to reduce brittleness.

A

Plasticizer

64
Q

• Rarelv used
• For histochemical, enzyme studies, and frozen sec.

A

Gelatin impregnation

65
Q

• Advantage: Water soluble (no dehydration and clearing needed)
• Disadvantage: may decay

A

Gelatin impregnation

66
Q

Gelatin impregnation

Advantages:
Disadvantages:

A

• Advantage: Water soluble (no dehydration and clearing needed)

• Disadvantage: may decay

67
Q

Gelatin impregnation

Tissue must be______ thick

A

<2-3mm thick

68
Q

Gelatin impregnation

_______ must be added to prevent molds and prevent further damage such as tissue decay

69
Q

Gelatin impregnation

METHODS OF GELATIN INFILTRATION

A

Wash out fixative

Put tissue in 10% gelatin with 1% phenol

20% gelatin with 1% phenol

Fresh 20% gelatin with 1% phenol

Cool in refrigerator

10% formalin

70
Q

Gelatin vs. frozen section

A

May fixation ang gelatin

71
Q

Composition of paraplast

A

Pure paraffin

Synthetic plastic polymer (dimethyl sulfoxide)

72
Q

Substitute for paraffin wax which is more elastic and resilient

73
Q

Substitute for paraffin wax which is less brittle and less compressible

74
Q

This allows easier handling and cutting of suitably thin sections without any damage or distortion to the tissue and its cellular components.

A

Impregnation

75
Q

is the simplest, most common and best embedding medium used for routine tissue processing.

A

Paraffin wax

76
Q

Paraffin wax

To decrease viscosity and improve infiltration of the tissue, technologists often increase the temperature to above…

A

60°C or 65°C.

77
Q

Manual Processing
At least_____ changes of wax are required at 15 minutes intervals in order to ensure complete removal of the clearing agent from the tissue.

78
Q

In automatic processing…only_____ changes of wax are required to remove the clearing agent and properly impregnate the specimen.

79
Q

The presence of any odor in the_____ during final paraffin wax bath indicates that the paraffin wax needs to be changed.

A

clearing agent

80
Q

involves wax impregnation under negative atmospheric pressure inside an embedding oven.

A

Vacuum Embedding

81
Q

Vacuum hastens the removal of air bubbles and clearing agent from the tissue block, thereby promoting a more rapid wax penetration of the tissue.

This technique is particularly recommended for urgent biopsies, for delicate tissues such as lung, brain, connective tissues, decalcified bones, eyes, spleen and central nervous system.

A

Vacuum embedding

82
Q

Vacuum infiltration requires a vacuum infiltrator or embedding oven, consisting of wax baths, fluid trap and vacuum gauge, to which a vacuum of up to_______ is applied using a water or mechanical pump

83
Q

The degree of the vacuum should not exceed….

84
Q

Of the three methods of paraffin wax impregnation, _____\ gives the fastest result

A

vacuum impregnation

85
Q

Fresh wax should be filtered before use in a wax oven at a temperature 2°C higher than its melting point.

Wax that has been trimmed away from the impregnated tissue may be melted and filtered for future use, with a coarse filter paper, e.g…

A

Green’s No. 904.

86
Q

When wax has been reused, some amount of water inevitably is mixed with it.

If excessive, this may impair the impregnating capacity of the medium and prevent formation of a good tissue block.

Water must therefore be removed by heating the wax to_____, thereby raising its melting point.

Paraffin wax may be used only_____, after which, fresh wax must be utilized.

A

100 -105°C

twice

87
Q

Paraplast

It is more elastic and resilient than paraffin wax thereby permitting large dense tissue blocks such as ____ and _____ to be cut easily with the same result as in double embedding

A

bones and brain

88
Q

is synthetic wax substitute similar to Paraplast with a melting point of 56-58°C (56-57°C)

89
Q

Sectioning of ester wax-impregnated tissues should be done on a heavy duty microtome (e.g.__________) due to the relative hardness of the wax.

A

sliding or sledge type microtome

90
Q

Water Soluble Waxes
are plastic polymers, mostly _____with melting points of 38-42°C or 45-56°C.

A

polyethylene glycols

91
Q

Due to its hygroscopic nature,_____ is very easily dissolved in water.
Hence care must be taken to avoid contact of the block with water or ice.

Tissue sections are very difficult to float out and mount due to its extreme solubility in water, dehydrating and clearing agents. Adding____ to water or using______ in water will reduce tissue distortion and promote flattening and “floating out” of sections.

A

Carbowax

soap

10% Polyethylene Glycol 900

92
Q

This is used mainly for preparing soft tissue sections of mixed consistency such as eyes and brain

93
Q

Disadvantages include inability to cut thin sections, storage of blocks in alcohol and speed of technique (which can take several weeks or months).

94
Q

is rarely used except when dehydration is to be avoided and when tissues are to be subjected to histochemical and enzyme studies.

A

Gelatin impregnation

95
Q

It is used as an embedding medium for delicate specimens and frozen tissue sections because it prevents fragmentation of tough and friable tissues when frozen sections are cut

96
Q

Tissues should not be more than_____ thick since gelatin-embedded specimens are harder to freeze than non-impregnated tissues

97
Q

In gelatin impregnation…The ___serves to prevent the growth of molds.

A

1 % phenol

Histopath LEC (37 decks)

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