IMPREGNATION AND EMBEDDING Flashcards by Janah Belaguin

___has a wide range of melting points which is important for use in the fifferent climactic regions of the world
Inexpensive
Provides quality sections
Can be used for most routine and special stains

Paraffin wax

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LOW MELTING POINT PARAFFIN – usually softer
HIGH MELTING POINT PARAFFIN – usually harder
Melting points range from ___ (commonly 56C)

40-70C

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Paraffin wax that contains plasticizers or other resin additives are commercially available
These mixtures create paraffin with the desired hardness for the tissues to be embedded
Additives have a high melting point than paraffin which makes the tissues brittle.
Examples:

Beeswax
Rubber
Ceresin
Plastic polymers
Diethylene glycol disteareate

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Sub for paraffin wax.
Is a mixture of highly purified paraffin and synthetic plastic polymers
Melting point: 56-57C
More elastic and resilient than paraffin wax thereby permitting large dense tissue blocks such as bones and brain to be easilt cut
Better ribboning of tissues and serial sectioning is allowed

PARAPLAST

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similar to paraplast with a melting point of 56-58C which is less pressible than paraplast

EMBEDDOL

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recommended for embedding eyes

BIOLOID

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paraffin plus rubber with the same property as paraplast

TISSUE MAT –

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Lower melting point (46-48C) but is harder than paraffin
Non water soluble but is soluble in 95% ethyl alcohol and other clearing agents
Three to four changes are needed
Cutting should be done with a sliding or sledge type microtome

ESTER WAX

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Mostly are polyethylene glycols with melting points of 38-42C or 45-56C

WATER SOLUBLE WAXES

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appears solid at room temperature
Does not need dehydration and clearing
Does not remove neutral wax
Good for enzymatic studies
Used in four changes : 70% (30mins)-90% (45 mins) and 100% (X2) (1H) at a temperature of 56C
Tissues are difficult to float out due to water miscibility (remedied by _ or _ )

CARBOWAX

soap or 10% polyethylene glycol 900

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Also known as COLLODION
Is a purified form of nitrocellulose soluble in many solvents suitable for specimens with large hollow cavities which tend to collapse, for hard and dense tissues such as bones and teeth and large tissue sections of the embryo
Supplied in:

CELLOIDIN

Thin (2%)
Medium (4%)
Thick (8%)

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Disadv.

Impregnation is very slow (days to week)
Very thin sections (less than 10u) are difficult to cut
Serial sections are different to prepare
Vapor of ether solvent is very flammable
Photomicrographs are difficult to obtain
Very volatile

CELLOIDIN

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Cellodin Methods

Recommended for bones, teeth, large brain sections, and whole organs

Recommended for the eyes

WET METHOD

DRY METHOD

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involves the enclosing of properly processed, correctly oriented specimens in a support medium that provides external support during microtomy

EMBEDDING

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consists of two L shaped strips of heavy brass or metal arranged on a flat metal plate

LEUCKHART’S EMBEDDING MOLD

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is made up of a series of interlocking plates resting on a flat metal base forming several compartments

COMPOUND EMBEDDING UNIT

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DISPOSABLE EMBEDDING MOLDS

Peel away
Plastic ice trays
Paper boats

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BLOCKING OUT MOLDS 4

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DISPOSABLE EMBEDDING MOLDS
PLASTIC EMBEDDING RINGS AND BASE MOLD
COMPOUND EMBEDDING UNIT
LEUCKHART’S EMBEDDING MOLD

Advantages:

Ease of use
Speed
Tissue and holder are firmly attached creating a single unit
Blocks filled immediately after sectioning
Permanent identification

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EMBEDDING CENTERS

EMBEDDING CENTERS Consists of three modules:

–allows the paraffin to solidify and ensures a small crystalline structure producing fewer artifacts when sectioning the tissue

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COLD PLATE
PARAFFIN DISPENSER
HEATED STORAGE AREA

COLD PLATE

Arteries, veins, fallopian tubes and vas deferens

Should be cut in cross sections of the lumen

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TUBULAR STRUCTURES

Cut in the plane at right angles to the surface

Oriented so that the epithelial surface is cut last minimizing compression and distortion of the epithelial layer

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SKIN, INTESTINE, GALLBLADDER, and OTHER EPITHELIAL BIOPSIES

Sections containing both transverse and longitudinal planes

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MUSCLE BIOPSIES

Oriented side by side with the epithelial surface facing the same directions

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MULTIPLE PIECES OF TISSUES

The process where tissues are first infiltrated with Celloidin and subsequently embedded in parrafin Used to facilitate cutting of large blocks of dense firm tissues like the brain

DOUBLE EMBEDDING METHOD

AUTOMATED TISSUE PROCESSING The basic principle for tissue processing requires the exchange of fluids using a series of solutions for a predetermined length of time in a controlled environment

No change for years | Recent advances now includes specialty MICROWAVE OVENS and the emergence of constant THROUGH-PUT processors

_ and _ were the first automated tissue processors used in histology laboratory.

The CAROUSEL TYPE PROCESSOR (tissue transfer) and the SELF CONTAINED FLUID EXCHANGE SYSTEMS

AUTOMATED TISSUE PROCESSORS | Makes use of an automatic tissue processing machine called

AUTOTECHNICON

The machine fixes, dehydrates, clears and infiltrates tissues, thereby decreasing the time and labor needed during the processing of tissues Impregnation is hastened by constant tissue agitation

AUTOTECHNICON

Permits the turning of platforms and easy access to beakers and wax baths. It makes use of 12 individual processing steps with 1 liter capacity glass beakers and two thermostatically controlled wax baths.

AUTOMATED TISSUE PROCESSORS

THE ___ controlled by electrical current moves the tissues from one reagent to another (by clock schedules)

TRANSFER ARM

Involves the wax impregnation under negative atmospheric pressure inside the embedding oven to hasten removal of air bubbles and clearing agent from the tissue block thereby promoting a more rapid wax penetration of tissues

VACUUM EMBEDDING

Time is reduced from 25-75% | Tissues are not over-exposed to heat - brittleness, shrinkage, and hardening of tissues is therefore prevented

VACUUM EMBEDDING

Hourly throughput of up to 120 specimens.

Tissue-Tek Xpress® Continuous Rapid Tissue Processor

This instrument uses an innovative, low-wattage microwave technology molecular-friendly reagents traditional vacuum infiltration techniques Specimen slides now make their way to the pathologist faster than ever, enabling same-day diagnosis and thus drastically reducing patient stay and anxiety.

Tissue-Tek Xpress® Continuous Rapid Tissue Processor

The Xpress® reagents preserve _ _ and _ in the paraffin block, eliminating the need to use fresh tissue for molecular studies.

DNA, RNA and proteins

The reagents provide excellent morphology, sharp nuclear detail and crisp cellular images equal to or better than conventional tissue processing. Compatible with special staining and immunohistochemistry methods, these reagents can often eliminate the need for antigen retrieval.

Tissue-Tek® Xpress® Continuous RapidTissue Processor

is the fifth generation of instruments for the enclosed automatic tissue processing of human and animal tissue specimens.

The Tissue-Tek® VIP® 5 Vacuum Infiltration Processor

20 user-defined processing programs for fixation, dehydrating, clearing and paraffin infiltration of tissue specimens.

The Tissue-Tek® VIP® 5 offers

provides more efficient processing using a redesigned control panel for easy monitoring of the system status and several preventive functions to protect against unexpected situations. Capacity of __ cassettes

The Tissue-Tek® VIP® Jr. 150

The system to fully automate your embedding No more opening of the cassette from grossing through sectioning Continuous loading of your blocks after processing A throughput of 120 blocks per hour, matching the capacity of the Tissue-Tek® Xpress® Continuous Rapid TissueProcessor

Tissue-Tek® AutoTEC® Automated Embedder

MAINTENANCE TIPS Any spillage or overflow should be wiped away immediately Accumulation of wax in any surface should be removed Temperature of the paraffin bath should be set to 3C above the melting point of the paraffin Timing should be checked when placing tissue cassettes in the processor, specially when delayed schedules are selected

Rarely used today There can be circumstances the tissue sample requiring the tissue sample to be manually processed, including Power failure and equipment malfunction Large tissue samples requiring more time that allocated by the automated processor Small biopsies, such as transplant specimens needing a rapid diagnosis

MANUAL TISSUE PROCESSING

IMPREGNATION AND EMBEDDING Flashcards

(43 cards)