Infiltration/Impregnation

Remove clearing agents

Fill up cavities and tissue spaces

Purpose of Infiltration/Impregnation

Paraffin Wax Infiltration

Routine tissue processing

Procedure is rapid

paraffin wax

Infiltrating agent of Paraffin Wax Infiltration

Serial, staining

Paraffin Wax Infiltration

Allows cutting of _______ sections

Compatible with _________procedures

hardening and brittleness

Paraffin Wax Infiltration

Prolonged process may cause

retention of clearing agent

Paraffin Wax Infiltration

Inadequate process may cause

Fats

Paraffin Wax Infiltration

NOT FOR tissues with ______

56 degC

Paraffin Wax Infiltration

Melting point for routine work

Manual

Automatic

Vacuum

Methods of Paraffin Wax Impregnation

Manual PWI

Carried out using Paraffin oven to keep the wax melted

55 - 60°C/2 - 5° above wax melting point

Temp of Manual PWI

4, 15

Manual PWI:
Requires __ changes of wax at _ minutes’ interval

Automatic PWI

Procedure that uses autotechnicon

4 steps (fixation to infiltration)

Autotechnican can only do ___ steps

2-3

Automatic PWI requires __ changes of wax

2-3

How many stations does Automatic PWI have

Tissue transfer type

Types of Automatic PWI:

tissue automatically transferred from one beaker/rgt to another; dip and dunk

Fluid transfer type/ enclosed

Types of Automatic PWI:

tissue stays in a chamber where the reagent in each step is pumped in and out

Vacuum PWI

What PWI uses Infiltration under negative atmospheric pressure inside the oven and is the fastest

Vacuum PWI

PWI Recommended for urgent biopsies, for specimens like lungs, brain, eyes, spleen, and CNS

Vacuum PWI

What PWI:
Removal of bubbles and clearing agent is rapid

Paraplast

Substitute for Paraffin Wax:
mixture of highly purified paraffin and synthetic plastin polymers

56–57°C

Melting point of Paraplast

Paraplast

Substitute for Paraffin Wax used for bones and brains

Substitute for Paraffin Wax:
similar to paraplast

Substitute for Paraffin Wax:
similar to paraplast

56–57°C

Melting point of Embeddol

Bioloid

Substitute for Paraffin Wax used for eye spx

Ester wax

Substitute for Paraffin Wax:

Harder than paraffin and requires heavy duty type of microtome e.g. sliding microtome

3-4 changes

How many changes are required for Ester wax

46–48°C

Melting point of Ester wax

46–48°C

Substitute for Paraffin Wax:
product of paraffin with rubber

Carbowax

Substitute for Paraffin Wax:
A water-soluble wax for enzyme histochemistry

Very hygroscopic

Collodion

Celloidin Infiltration is also called?

Celloidin

Fills up cavities
A purified form of nitrocellulose

Celloidin Infiltration

For tissues with very large cavities and spaces that tends to collapse

2
4.8
equal
ether
alcohol
heat

Celloidin Infiltration

In , _% solutions dissolved in _____parts of _____and _______
No ____applied; Very slow, thin sections

photomicrography

Celloidin Infiltration
Sections for ____________are all difficult to obtain thin sections

2-4%

Concentration of Thin Celloidin

4-6%

Concentration of Medium Celloidin

8-12%

Concentration of Thick Celloidin

5-7 days

Duration of Thin Celloidin

5-7 days

Duration of Medium Celloidin

3-5 days

Duration of Thick Celloidin

It becomes more volatile

What happens when you increase the concentration of Celloidin during Infiltration hence making it not recommended for speeding up the process

Same

Dry and Wet Celloidin have the ___ procedure

Wet Celloidin

Celloidin used for Brain, bones, whole organs

70% alcohol

Reagent used to store Wet Celloidin

Dry Celloidin

Celloidin used for Eye specimen

Gilson’s Mixture

Reagent used to store Wet Celloidin

Nitrocellulose Method

Also called Low Viscosity Nitrocellulose

Nitrocellulose Method

Another form of celloidin soluble in equal concentrations of ether and alcohol

Highly explosive

Equal
ether
alcohol

Explosive

Nitrocellulose Method

Another form of celloidin soluble in _____ concentrations of _____and ______

Highly ______

Nitrocellulose Method

Produces harder tissue blocks and allows cutting of thinner sections

Tissues have tendency to crack

Addition of Oleum ricini or castor oil

Nitrocellulose Method

Tissues have tendency to crack
What remedy?

Gelatin Infiltration

Rarely used infiltration method

20-30x the volume of specimen

Required volume of Gelatin infiltration

Gelatin Infiltration

Infiltration used in enzyme studies

Soluble
dehydration
clearing

Gelatin Infiltration is water ______ which eliminates _______ and _______

2-3mm/ 3-4mm thick

Thickness of tissue for Gelatin Infiltration

1% phenol

What is added to Felatin Infiltration to prevent molds

10% gelatin and 1% phenol for 24hrs

Process of Gelatin Infiltration
Number 1

20% gelatin and 1% phenol for 12hrs

Process of Gelatin Infiltration
Number 2

20% gelatin and 1% phenol

Process of Gelatin Infiltration
Number 3

Embed

Process of Gelatin Infiltration
Number 4

Formalin

Process of Gelatin Infiltration
Number 5

Hardening

Process of Gelatin Infiltration
Number 6