Two general methods of dehydration

• Graded dehydration series: The gradual replacement of water with an

organic solvent by transferring the tissue through a series of increasingly
concentrated solutions like ethanol, acetone etc.
• Rapid dehydration series: It is rapid dehydration method using an organic
solvent. It is less common. In this method tissue water is replaced rapidly,
without the surface tension damage that can be associated with alcohols or
acetone. It is limited to plastic embedding methods.
Dioxane 
Also known as diethylene dioxide, this is a colourless, flammable liquid
that produces less shrinkage and hardening than with ethanol.
• The liquid is miscible with water, most organic solvents and paraffin
wax and is excellent for tissues that have been excessively hardened
by conventional processing.
• Dioxane has a rapid but gentle action and tissues are able to remain
in it for long periods without harm.
• Benzene

Benzene is more gentle and rapid than xylene and toluene and is probably
the best transition solvent However, its toxicity and carcinogenicity has
preclude its use in histology.

Butyl acetate

This is used as a xylene substitute and nitrocellulose solvent.

Carbon tetrachloride

Because of its high toxicity it is now rarely used in histology.

• Chloroform

This is an expensive, volatile but slowly penetrating solvent. It causes less

brittleness than xylene and is often used on dense tissues such as uterus.
However, it attacks some plastics and sealants so is not generally
recommended for enclosed processors.

Limonene (d-limonene)

This is derived from citrus fruit and is a component of various proprietary

blends of transition solvents such as Histoclear and Citroclear marketed as
xylene substitutes. It is less viscous than cedarwood oil and is similar to the
esters in clearing action and in elimination from wax. Limonene may cause
allergic skin reactions.
• Trichloroethane

This is commonly used as a xylene substitute and is a component of agents

such as Inhibisol and CNP30. These solvents are stable to light but tend to
slowly liberate hydrochloric acid on contact with water. Because of their
high volatility, members of this group may achieve and exceed maximum
allowable concentrations in poorly ventilated laboratories far more rapidly
than xylene under the same conditions.