DEHYDRATION

Dehydration
-- The process of removing intercellular and
extracellular water from the tissue following
fixation and prior to wax impregnation

Dehydrating Agents
- solutions utilized in the dehydration process.
Characteristics of an Ideal Dehydrating
Solution
1.It should dehydrate rapidly without producing
considerable shrinkage or distortion of tissues
2.It should not evaporate fast
3.It should be able to dehydrate even fatty tissues
4.It should not harden tissue excessively
5.It should not remove stains
6.It should not be toxic to the body
7.It should not be a fire hazard
Commonly used dehydrating
agents:
1.Alcohol (most common)
2.Acetone
3.Dioxane 4 - cellusolve
4.Triethyl phosphate
5.Tetrahydrofuran
Alcohol
•Ethyl alcohol (ethanol)
the alcohol recommended for routine dehydration
of tissues
- clear, colorless, flammable fluid
- considered to be the best dehydrating agent
because it is fast - acting, mixes with water and many
organic solvents, and penetrates tissue easily
- not poisonous and not very expensive
Methyl Alcohol
- toxic dehydrating agent
- primarily employed for blood and tissue films and for
smear preparation
Butyl Alcohol
-utilized in plant and animal micro-techniques
-a slow dehydrating agent, producing less shrinkage
and hardening than ethyl alcohol
- recommended for tissues which do not require rapid
processing
 10% Zenker or Bouin’s Susa, Carnoy Flemming’s
Formol Helly’s Fluid or Formol- Fluid
Saline Sublimate

Running Water 1-12 1-12

Alcohol 30 % 1-6 1/2- 3
Alcohol 50% 1-6 1/2-3
Alcohol 70% 3-12 1-6 3-12 1/2 - 3
Alcohol 90% 3-12 1-6 3-12 1-6 1-3
Absolute 1-2 1-2 1-2 1-2 1-2
Alcohol
Absolute 1-2 1-2 1-2 1-2 1-2
Alcohol
Absolute 1-2 1-2 1-2 1-2 1-2
Alcohol
 Acetone
- cheap, rapid acting dehydrating agent utilized for most urgent
biopsies which dehydrates in 1/2 to 2hrs
- clear, colorless fluid that mixes with water, ethanol and most
organic solvents
- more miscible with epoxy resins than alcohol but is highly
flammable and requires considerable care in handling
-rapid in action but penetrates tissue poorly and causes brittleness
in tissues that are placed in acetone for prolonged period of time
- not recommended for routine dehydration purposes
Dioxane (Diethylene Dioxide)
- excellent dehydrating and clearing agent readily
miscible in water, melted paraffin, alcohol and xylol
- produces less shrinkage compare to alcohol
dehydration
- expensive and extremely dangerous
Cellosolve (Ethylene glycol monoethyl
ether)
• Cellosolve - dehydrates rapidly
- toxic by inhalation, skin contact, and ingestion
- use propylene-based glycol ethers instead
Triethyl Phosphate
- removes water readily and produces very little
distortion and hardening of tissue
- soluble in alcohol, water, ether, benzene, chloroform,
acetone and xylene
Tetrahydrofuran THF
- both dehydrates and clears tissues since it is miscible
in both water and paraffin
-toxic if ingested or inhaled
- prolonged exposure (6months) may cause
conjunctival irritation