HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES - LECTURE

LESSON 5: CLEARING
MIDTERMS | A.Y. 2022-2023 | PROF. DOREN VENUS OTOD

OUTLINE IV. TYPES OF CLEARING AGENT

I. Introduction to Clearing
II. Purpose of Clearing A. ADVANTAGE AND DISADVANTAGES OF ALCOHOL
III. Characteristics of Good Clearing Agent AGENTS
IV. Types of Clearing Agents
A. Advantages and Disadvantages of Alcohol ALCOHOLS
Clearing Agents AGENT ADVANTAGES DISADVANTAGES
B. Common Clearing Agents
C. Others Clearing Agents XYLENE ● rapid, suitable ● Highly
30mins to for urgent flammable
1 hr biopsies ● > 3 hours,
CLEARING ● Makes the tse makes tse hard
transparent ● Not suitable for
I. INTRODUCTION ● Miscible both in nervous tissues
alcohol and and lymph
FIXATION ➨ DECALIFICATION ➨ DEHYDRATION ➨ paraffin nodes
CLEARING (DE-ALCOHOLIZATION) ● Cheap
TOLUENE ● Miscible both in ● Slower than
1hr to 2hrs alcohol and xylene
“In the process of Clearing, we can also term this one as paraffin ● More
de-alcholization because we are removing alcohol, which ● Does not make expensive
is the commonly used dehydrating agent during the process tse hard and
of dehydration.” brittle
● Not
II. PURPOSE OF CLEARING carcinogenic
BENZENE ● For URGENT ● Carcinogenic
15mins to biopsies ● It may lead to
1. Removing dehydrant from tissue, and replacing
1hr ● Fastest aplastic anemia
it with a fluid miscible to both dehydrant and clearing agent
embedding agent.
2. Makes tissues “translucent” or transparent, B. COMMON CLEARING AGENTS
hence the term clearing.
3. Most are flammable fluids and have low boiling CHLOROFORM (6-24 hours)
points ● Tough tissues and large specimens
4. Excessive clearing may cause brittleness ○ Skin, fibroid and decalcified tissues
(requires longer processing time)
● The main purpose of performing clearing is to ● Toxic to liver (prolong inhalation)
create a bridge between the process of
dehydration and infiltration. CEDARWOOD OIL (2-3 days)
● It is because our paraffin wax is not miscible or ● For smooth muscle, CNS
compatible with our alcohol, so we need to (requires deeper penetration)
remove the alcohol thru the process of clearing, ● Milky on prolonged storage
and preparing our tissue for the next process,
which is the infiltration. ANILINE OIL
● Prolonged exposure to clearing agents may result ● For insects, embryos, and delicate tissues
to some respiratory problems.
CLOVE OIL
III. CHARACTERISTICS OF GOOD CLEARING AGENT ● Minimum shrinkage of tissues
● Slow, expensive and difficult to use
● It should be miscible with alcohol. (not usually used in the laboratory)
● It should be miscible with paraffin wax.
● It should not produce tissue shrinkage. CARBON TETRACHLORIDE
● It should make tissue transparent. ● Tough tissues and large specimens
● It should not dissolve aniline dyes. (same tse of choice to chloroform)
● It should not evaporate quickly in water bath ○ Skin, fibroid and decalcified tissues
● Similar to chloroform but cheaper
● Dangerous to inhale on prolonged exposure

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TETRAHYDRFURAN
● Nontoxic but with offensive odor and should be
used in a well-ventilated room
● This considered superior among other agents
since it can process both DEHYDRATION AND
CLEARING thus shorter processing time
● It has an offensive, ethereal odor so the room must
be well-ventilated

DIOXANE
● Causes greater shrinkage than xylene
● Dangerous, toxic to liver

C. OTHER CLEARING AGENTS

OTHERS
GUM SYRUP & ● Used when clearing directly from
GLYCERIN water ( as in a frozen section)
OIL OF ● For skin and smooth muscle
BERGAMOT
OIL OF ● For skin
ORIGANUM
OIL OF ● Artificial oil, for delicate tissues
WINTERGREEN
CARBON ● For smooth muscle, has foul
DISULFIDE odor
CARBOL ● For friable tissues
XYLENE
TERPINEOL ● For delicate tissues like eyes
PHENOL ● For smooth muscles
HIGH TEST ● Excellent clearing agent
AVIATION
LEAD-FREE
GASOLINE

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