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Clearing Trans

The document outlines the process of fresh tissue specimen preparation for examination, focusing on clearing agents used to remove dehydrating agents and replace them with fluids compatible with embedding media. It details various clearing agents such as xylene, toluene, and benzene, including their properties, recommended uses, and potential drawbacks. Additionally, it discusses the importance of proper dehydration to maintain tissue transparency and refractive index.

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Mark Villapando
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31 views2 pages

Clearing Trans

The document outlines the process of fresh tissue specimen preparation for examination, focusing on clearing agents used to remove dehydrating agents and replace them with fluids compatible with embedding media. It details various clearing agents such as xylene, toluene, and benzene, including their properties, recommended uses, and potential drawbacks. Additionally, it discusses the importance of proper dehydration to maintain tissue transparency and refractive index.

Uploaded by

Mark Villapando
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

FRESH TISSUE SPECIME N PREPARATION FOR EX AMINATION

CLEARING

 AKA: Dealcoholization CHLOROFORM


 Removal of dehydrating agent and replacement of a fluid that is
miscible with both dehydration agent and embedding medium  Clearing: 6-24 hours
 Translucent — increased refractive index  Slower in action than xylol
 Can be used for tissue blocking
ROUTINE TISSUE PROCESSING  Recommended for tough tissue
 Tissue do not become translucent
 CLEARING AGENTS
 Cause less brittleness
o Remove alcohol
 Tissue blocking up to 1cm
o Flammable liquids
 Tissue tend to float
o Low boiling points
 Hepatotoxic
 DURATION OF CLEARING
o Over clearing may cause tissue brittleness
ANILINE OIL
COMMONLY USED CLEARI NG AGENTS
 Not for routine purposes
 Recommended for embryos, insects, and very delicate
XYLENE tissues
 Other name: Xylol
 Colorless clearing agent CLOVE OIL
 Most common routine procedure
 Rapid clearing time: 15-30 minutes to 1 hour  Minimum shrinkage
 Used in Celloidin section  Adulterated —> Easily contaminated
 Tissue collection thickness: <5mm  Recommended for skin and smooth muscles
 Milky when completely dehydrated  Not for routine purposes

TLOUENE CEDARWOOD OIL


 Other name: Toluol  Recommended for CNS tissue and cytology
 Clearing: 1-2 hours
o Smooth muscle and skin
 Alternative for xylene and benzene
 Require change (twice) of clearing agent
 If left in toluene for 1 day — will not be excessively hard or
brittle  No tissue distortion even if tissue is left indefinitely
 Not carcinogenic  Disadvantages:
 Toxic upon prolonged exposure o Very slow
 2-3 days
BENZENE o Very expensive
o Tend to become milky upon prolonged storage
 Clearing: 15-60 minutes
 Doesn’t make tissues hard and brittle; minimum shrinkage
 Recommended for urgent biopsy and routine purposes CARBON TETRACHLORIDE
 Highly toxic
 Similar with Chloroform
o Can cause aplastic anemia
 Cheaper
 It can cause tissue hardening
METHYL BENZOATE AND METHYL SALICYLATE
 Toxic if prolonged exposure
 Slow acting
 Double embedding techniques

VILLAPANDO, MARK ANDREW OBEJERA 1


HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES

OTHER CLEARING AGENTS PROCEDURE

 Clear dehydrated tissue in changes of benzene — 1 hour


OIL OF BERGAMOT each
 FOR SKIN AND SMOOTH MUSCLE  Note: Insufficient dehydration will turn clearing agent
milky

OIL OF ORIGANUM
 For skin

OIL OF WINTERGREEN
 Artificial oil
 For delicate tissues

CARBON DISULFIDE
 For smooth muscle
 Foul odor

CARBOL XYLENE
 For friable tissues

TERPINEOL
 For delicate materials like eyes

PHENOL
 For smooth muscles

HIGH TEST AVIATION L EAD FREE GASOLINE


 Excellent clearing agent

LABORATORY

DEHYDRATION

 Other name: Dealcoholization


 Removing the alcohol in tissue
 Why remove the alcohol?
o Many substances have property of making tissue
transparent
o It affects their high refractive index
o Refractive index = Tissue

MATERIALS AND REAGENTS

 Bottles
 Benzene

VILLAPANDO, MARK ANDREW OBEJERA 2

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