[LAB] Unit 5: Impregnation & Embedding Flashcards by kehliess dumdum

Process of saturating the tissue with a medium, usually liquid paraffin, to permeate or fill up the natural cavities, spaces, and interstices of the tissue.

Impregnation

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Impregnation is the process of ______ing the tissue with a medium, usually liquid paraffin, to permeate or fill up the natural cavities, spaces, and interstices of the tissue.

Impregnation is the process of SATURATING the tissue with a medium, usually liquid paraffin, to permeate or fill up the natural cavities, spaces, and interstices of the tissue.

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These professionals determine what tissue to use/process

Pathologists

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(T/F)
Histotechnologists cut the tissue to be used

FALSE

Pathologists are the ones to cut the tissue to be used

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What does FFPE stand for?

Formalin Fixed Paraffin Embedded

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A suitable _________ ______ is filled with the molten wax, the tissue is placed in it and oriented so it is sectioned in the proper plane.

Embedding mold

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(T/F)
A variety of molds can be used depending on the
technician’s preference

True

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What is the output of embedding?

Tissue block

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(T/F)
Cooling the tissue block rapidly would avoid crystals

True

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EMBEDDING TECHNIQUES

(1.)
Transfer the tissue with _____ forceps to a small container of freshly ______ paraffin

Transfer the tissue with warm forceps to a small container of freshly melted paraffin

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EMBEDDING TECHNIQUES

(T/F)
Tips of forceps are heated in an alcohol lamp or a hotplate

FALSE.

Tips of forceps are heated in an alcohol lamp or in a forceps warmer

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EMBEDDING TECHNIQUES

Forceps tip should be hot enough so paraffin does not ______ , but not so hot as to cause paraffin to ______.

Forceps tip should be hot enough so paraffin does not solidify , but not so hot as to cause paraffin to smoke.

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EMBEDDING TECHNIQUES

(2.)
Fill the bottom of the mold with a small amount of _______.

The depth of the mold should be at least _____the thickness of the tissue.

Fill the bottom of the mold with a small amount of **paraffin **

The depth of the mold should be at least TWICE the thickness of the tissue.

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EMBEDDING TECHNIQUES

(3.)
Pick up tissue, and place into the mold. Manipulation of the tissue in the mold must be ______, so paraffin does not begin to harden.

Pick up tissue, and place into the mold. Manipulation of the tissue in the mold must be quick, so paraffin does not begin to harden.

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(T/F)
The tissue should be placed on the bottom part of the mold, so that when the tissue block is retrieved, the cutting face will not be exposed when sectioning

FALSE.
The tissue should be placed on the bottom part of the mold, so that when the tissue block is retrieved, the cutting face will be easily exposed when sectioning

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(T/F)
Place the tissue cassette on the metal mold to act as a cover

True

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EMBEDDING TECHNIQUES

(4.)
After the tissue is in the mold, fill the mold entirely with the paraffin. As the paraffin begins to harden, insert a ____ ______ _____; the label should not go down to the bottom of the paraffin.

(4.)

After the tissue is in the mold, fill the mold entirely with the paraffin. As the paraffin begins to harden, insert a code number label; the label should not go down to the bottom of the paraffin.

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EMBEDDING TECHNIQUES

(T/F)
the label should go down to the bottom of the paraffin.

False

the label should NOT go down to the bottom of the paraffin.

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EMBEDDING TECHNIQUES

This refers to how the tissue is placed in the wax
as it hardens

Tissue Orientation

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EMBEDDING TECHNIQUES

(T/F)
When embedding a skin specimen, its different layers should be seen in the mold.

True

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EMBEDDING TECHNIQUES

After filling the mold entirely with the paraffin wax, place immediately on ____ ___________ ________ to solidify

After filling the mold entirely with the paraffin wax, place immediately on cold orientation platform to solidify

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EMBEDDING TECHNIQUES

(5.)
Allow the surface of the paraffin block to harden, then immerse the mold into a _______, _____ _ _ ̊C water bath for about _ _ - _ _ minutes to hasten solidification of the paraffin.

Allow the surface of the paraffin block to harden, then immerse the mold into a shallow, cool (10 ̊C) water bath for about 10-15 minutes to hasten solidification of the paraffin.

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EMBEDDING TECHNIQUES

(7.)
When paraffin is completely _____, _____ it from the mold.

When paraffin is completely hardened, remove it from the mold.

*Do not immediately remove the tissue block from
the mold

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COOLING TEMPERATURE

___ ̊C temperature prevents cracking of the tissue block

10 ̊C temperature prevents cracking of the tissue block

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# COOLING TEMPERATURE 10 ̊C temperature prevents ______ of the tissue block

10 ̊C temperature prevents **cracking** of the tissue block

# COOLING TEMPERATURE If paraffin is properly cooled, the crystals of paraffin are ______ and ________ with each other.

If paraffin is properly cooled, the crystals of paraffin are **small** and **contiguous** with each other.

# COOLING TEMPERATURE (T/F) If properly cooled, the paraffin will appear turbid and homogeneous and there is no layering of the paraffin.

False If properly cooled, the paraffin will appear **clear** and homogeneous and there is no layering of the paraffin.

# COOLING TEMPERATURE What are the characteristics of a properly cooled tissue block?

-Clear -Homogenous -No layering of paraffin

# COOLING TEMPERATURE (T/F) Paraffin demonstrating these conditions (clear, homogenous, no layering) are not ideal for sectioning.

False. Paraffin demonstrating these conditions is BEST for sectioning

This infiltrating medium is rapidly converted from solid to liquid form on heating

Paraffin

This medium permeates the tissue in a liquid state

Paraffin

Paraffin permeates the tissue in what state

liquid state

(T/F) Paraffin solidifies relatively quickly on cooling

True

(T/F) Paraffin becomes fluid on heating to a temperature which may damage the tissue

False. temperature which will **not damage the tissue**

When the paraffin solidifies, it becomes firm enough to section at what temperature?

Room temperature

# Paraffin Time of infiltration and subsequent embedding are relatively short for ______ pieces of tissue.

small

# Paraffin Thin sections can be cut with the _______ microtome and sections will adhere to each other to form a _____.

rotary microtome; ribbon

# Paraffin (T/F) Tissue once infiltrated and embedded can be stored in a **dry condition** **indefinitely** without damage to the tissue.

True

# Paraffin What microtome is used for paraffin?

Rotary microtome

# Paraffin (T/F) You can wrap the paraffin in a tissue paper

True

Substitutes for Paraffin Wax

1. Paraplast 2. FIbrowax 3. Bioloid 4. Embeddol 5. Ester Wax 6. Water Soluble Waxes 7. Dimethylsulfoxide | PWD BEEF

This substitute for paraffin wax is a mixture of **highly purified paraffin** and **synthetic plastic polymers**

Paraplast | para: highly purified **para**ffin; plast: synthetic **plast**ic polymer

This substitute for paraffin wax is **less brittle** and **less compressible** than Paraplast

Embeddol

Embeddol is less ____ and less _____ than paraplast

Less brittle and less compressible

# Embeddol This is a **semisynthetic wax** recommended for embedding **eyes**

Myeloid

# Embeddol Myeloid is a _______ wax recommended for embedding ____

semisynthetic wax; eyes

This is a highly purified paraffin wax with DMSO (dimethylsulfoxide)

Ester wax

Ester wax is a _______ ________ paraffin wax with _______.

highly purified ; DMSO | DMSO = Dimethylsulfoxide

This wax is elastic and resilient

Ester wax

This wax has a lower melting point (46-48°C) but is harder than paraffin.

Ester wax

What is the melting point of ester wax?

46-48°C

Ester wax is **not** soluble in water, but is soluble in _ _ % ______ ________ and other clearing agents

95% Ethyl Alcohol

This wax is **not** soluble in water

Ester wax

These waxes have melting points of 38-42°C or 45-56°C

Water soluble waxes

Water soluble waxes have melting points of ?

38-42°C or 45-56°C

# Water soluble waxes this water soluble wax is a polyethylene glycol

Carbawax

this water soluble wax is suitable for many **enzyme histochemical studies**

Carbawax

In this water soluble wax, **cytologic details** are **excellently preserved**

Carbawax

Carbawax is a p________ g_____

polyethylene glycol

Carbawax excellently preserves _________ ______

cytological details | (C=C) Carbawax = Cytologic

# DISADVANTAGES OF PARAFFIN When does the distortion of the histology of the tissue due to shrinkage usually occur?

When the sections are being attached to glass slides **(paraffin artifact)**

# DISADVANTAGES OF PARAFFIN (T/F) Sectioning of paraffin difficult at low temperatures

False. Sectioning of paraffin difficult at HIGH temperatures | because it will lose its rigidity

# DISADVANTAGES OF PARAFFIN (T/F) Time for infiltration of large blocks of tissue is excessive.

True

# DISADVANTAGES OF PARAFFIN (T/F) Paraffin shrinks after fixation

True | According to SIr

this infiltrating medium is an amorphous, slightly yellowish substance

Celloidin

This embedding medium is a purified form of collodion or nitro-cellulose

Celloidin

Celloidin is a purified form of _______ or ____-_______

Collodion or Nitro-cellulose

This embedding medium is for **hard tissue specimens**

Celloidin

What microtome is used for celloidin?

Sliding microtome

(T/F) Celloidin requires minimal heat

False. Celloidin does NOT require heat

(T/F) Celloidin has a robbery consistency

False. rubbery consistency

(T/F) Celloidin provides **minimal distortion** of specimen

True

(T/F) Celloidin is difficult to cut into thin sections

True

(T/F) With celloidin, serial sections are difficult to prepare

True

(T/F) Using celloidin is a fast process

FALSE. slow process

Celloidin blocks and sections must be stored in _ _ % __________ otherwise they become discolored, dry, and shrunken

70% alcohol

Celloidin blocks and sections must be stored in 70% alcohol otherwise they become ______, _______, and ________.

Discolored, dry, and shrunken

(T/F) Celloidin blocks and sections not stored in 70% alcohol = yellowish and shrunks

True

Which is more used? Dry celloidin method or Wet celloidin method?

Wet celloidin method

What does VCN stand for?

Low Viscosity Nitrocellulose

The low viscosity of LVN allows for ______ concentration to be used

higher

(T/F) LVN has greater speed of impregnation

True(

LVN's final block is harder, allowing _______ sections to be cut a. Thinner b. Thicker

a. thinner | The harder the block = easier to cut thinner sections ## Footnote Softer block = harder to cut thin sections

Which embedding medium has greater water tolerance? LVN or Celloidin?

Low Viscosity Nitrocellulose

(T/F) LVN has the tendency to crack down during handling and staining ## Footnote W

True

LVN has the tendency to crack down during handling and staining. What is used to minimize this tendency?

**0.5% oleum ricini** (Castor Oil)

Which embedding medium is HIGHLY EXPLOSIVE

LVN | bothmnemonic: TNT = LVN (both three letters)

In DOUBLE EMBEDDING, tissue is first impregnated with ________, and subsequently blocked in __________ ___

First: celloidin Subsequently: paraffin wax

(T/F) Double embedding is used when dealing with soft tissues

FALSE. HARD Tissues

Double embedding is used for __________ of the morphological appearance of the tissue

maintenance

Double embedding is used for maintenance of the __________ of the tissue

morphological appearance

(T/F) Serial sections are easily prepared with double embedding

True

With double embedding, extra degree of _______ is given when cutting hard tissues

Resilience

main use is in double embedding technique with **ester wax** or **paraffin wax**

Agar

cohesive agent for multiple fragments of friable tissue

Agar

It has a lower melting point that agar

Gelatin

Main use is in the production of whole organ sections

Gelatin

Which of the following are used for friable tissues: a. Gelatin b. Agar c. Both d. None of the above

c. Both

Water-soluble waxes can be embedded directly from ______. However, it is RESTRICTED due to the ______ diffusion currents which can lead to fragmentation of the section

Water ; Violent

What happens when you embed water-soluble waxes directly from water?

**diffusion currents** which can lead to **fragmentation** of the section

Give the three plastic embedding mediums

1. Epoxy 2. Polyester 3. Acrylic

This plastic embedding medium reduces antigenicity, is toxic, and damages tissue

Epoxy

Give the three epoxies used as an embedding medium

* bisphenol A (Araldite) * glycerol (Epon) * cyclohexene dioxide (Spurr) | BGC

This plastic embedding medium is NOT often used

Polyester

This plastic embedding medium is used extensively for l**ight microscopy**

Acrylic

Types of Acrylic

* MMA (Methyl Methacrylate) * GMA ( Glycidyl Methacrylate)

# MOLDS FOR EMBEDDING molds for routine work and are widely used

Leuckhart's Embedding Mold

Leuckhart's embedding mold consists of how many L-shaped pieces of metal?

2 L-shaped pieces of metal | L-shaped = Leuckhart

Leuckhart's embedding mold is arranged on a _____ metal plate to form a mold of desired size

glass

# Leuckhart's embedding mold (T/F) Any size of the block can be made and size of tissue blocks are resizable

True

This embedding mold consists of a **series of interlocking plates** resting on a flat metal base, forming **several compartments**

Compound Embedding Unit

This embedding mold has the advantage of embedding more specimens at a time

Compound Embedding Unit

This embedding mold is used in positioning histological tissues accurately in base molds.

Plastic Embedding Ring and Base Model

This embedding mold is compatible with most commonly-used processing and storage systems

Plastic Embedding Ring and Base Mold

In plastic embedding ring, _____ are precision-molded from premium-grade, chemically-inert, high impact polystyrene for dimensional rigidity and sturdiness.

Rings

(T/F) Base molds are like metal molds

True

This embedding mold allows tissue block to solidify and cool both sides of mold para lumabas yung block | ?

POP-OUT EMBEDDING MOLDS

Disposable embedding molds are ____-_-___

peel-a-way

_________: once the wax has solidified, the plastic walls are peeled off one at a time, giving perfect blocks that **require no trimming**.

peel-a-way (Disposable embedding mold)

# Dis (T/F) Once the disposable embedding molds are peeled off, they can be placed directly in the chuck of the microtome

True

# Molds for embedding It is made from thick paper or cardboard paper

Paper boat

# Molds for embedding It is cheap to make and allow blocks to be stored without being removed

Paper Boat

# Molds for embedding this type of paper is used para hindi dumikit yung paper sa block

glossy magazine

# Molds for embedding It provide easy and accurate identification of specimens, thereby avoiding confusion and interchange of tissue blocks

Paper boat

# Molds for embedding blocks are easily removed by flexing the plastic trays and by smearing the inside of the mold with glycerin or liquid paraffin

Plastic ice tray

# Molds for embedding convenient molds for **busy routine laboratory**, one block being embedded in each compartment

Plastic ice tray

What is smeared inside the mold of the plastic ice tray to be easily removed by flexing the plastic tray?

* Glycerin * Liquid paraffin

# Molds for embedding For busy laboratories (multiple blocks can be made from one tray)

Plastic ice tray

This is ideal for embedding fragmentary biopsies

Watch Glasses

(T/F) Watch glasses are not essential to smear them with glycerin

True

This is used for embedding tissue intended for EM microscopy

Methylcrylate plastic resin (EPON resin)

(T/F) Blocks are hard to remove from watch glasses

True

Used for small fragments which have been processed (e.g. **Bone marrow**) which concentrates them **without** the damage caused by orientation with forceps

Test tubes

Where in the tube do the small fragments concentrate?

Bottom

(T/F) You need to break the tubes in order to obtain the embedded specimen

True

# Pre-Test (T/F) Cellosolve is an embedding medium

FALSE | Celloidin

Match the corresponding guides on how to embed 1. Tubular tissue 2. Skin 3. Endometrial curetting 4. Long tissue 5. Intestine 6. Membrane A. all layers should come O. all layers should come L. all layers in transverse section N. swiss S. keep diagonally C. keep in center

# Pre-Test (T/F) After embedding, 10 ̊C temperature prevents cracking of the tissue block

True

# Pre-Test (T/F) Test tubes is one of possible embedding molds

True

# Post-Test (T/F) Low Viscosity Nitrocellulose is highly explosive

True

# Post-Test (T/F) Blocks are easily removed by smearing the inside with glycerin

True

# Post-Test (T/F) Tissues should not reach the edges during embedding

True