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The document discusses various tissue processing and embedding techniques for microscopy including celloidin processing, plastics, gelatin processing and orientation. Celloidin processing involves embedding specimens in celloidin solution and can be done via wet or dry methods. Plastics like epoxy and acrylic provide superior results for light microscopy. Gelatin processing is used for histochemical and enzyme studies as gelatin is water-soluble and has a low melting point. The document also covers orientation of tissues and different types of embedding molds.

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Marie Llanes
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52 views2 pages

Other Media

The document discusses various tissue processing and embedding techniques for microscopy including celloidin processing, plastics, gelatin processing and orientation. Celloidin processing involves embedding specimens in celloidin solution and can be done via wet or dry methods. Plastics like epoxy and acrylic provide superior results for light microscopy. Gelatin processing is used for histochemical and enzyme studies as gelatin is water-soluble and has a low melting point. The document also covers orientation of tissues and different types of embedding molds.

Uploaded by

Marie Llanes
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

CELLOIDIN PROCESSING ❖ Same with wet method EXCEPT for the following steps

✔ Purified pyroxylin nitrocellulose ✔ Gilson’s mixture (chloroform + cedarwood oil) is


✔ Suitable for specimens containing large cavities or hollow added to the fresh thick celloidin during embedding
spaces which tend to collapse (eyes) & for larger embryos ✔ Storage in alcohol is contraindicated
✔ Available in thin (2%), medium (4%) and thick (8%) solutions
✔ DIS: Tissues cannot be cut as thin as they are with paraffin PLASTICS
wax ❖ Provided superior results for light microscopic studies
✔ ADV: Causes much less shrinkage & distortion; slow process especially of hard tissues and samples for high resolution
With rubbery consistency – w/o distortion microscopy
Does not require heat
❖ Low Viscosity Nitrocellulose TYPES OF PLASTIC MEDIA
Soluble in equal concentrations of ether and alcohol 1. EPOXY (epoxy plastic, catalyst & accelerator)
ADVANTAGES: Araldite base (bisphenol) - slowest
✔ Can be used in higher concentrations Glycerol base (epon)
✔ Produces harder tissue blocks Cyclohexene Dioxide (spurr) - fastest
DISADVANTAGES: Disadvantages:
o Cracking of tissue sections and Reduce antigenicity
chrome-mordanted tissues may crumble Sensitization on skin contact and inhalation
REMEDY: add plasticizers like Castor oil, Contains toxic components (eg. Vinylcyclohexane
Oleum ricini dioxide)
o More explosive than celloidin 2. POLYESTER
o Cannot be stored 3. ACRYLIC
For high resolution LM
WET METHOD Glycol Methacrylate (GMA): valued for its hydrophilic
❖ for bones, teeth large brain sections & whole organs nature
❖ Fixation ​ ​ Dehydration ​ ​ Equal parts ether and alcohol Methyl Methacrylate (MMA): valued for its hardness
(12-24 hours) ​ ​ Thin celloidin: 2-4 % (5-7 days) ​ ​ Medium Benzoyl Peroxide​ ​– added as catalyst
celloidin: 4-6 % (5-7 days) ​ ​ Thick celloidin: 8-12 % (3-5 days) acts as an active site (polymerization of acrylics)
​ Embedding: fresh thick celloidin in a jar or desiccator
(fingerprint no longer leaves a mark on block surface) ​
Storage: 70-80% alcohol
GELATIN PROCESSING
DRY METHOD ✔ Histochemical and enzyme studies
✔ Delicate specimens
✔ Frozen sections
✔ Water-soluble
✔ Low MP and does not overharden
❖ Fixation ​ ​ washing ​ ​ 10 % (24 hours) ​ ​ 20 % (12
hours) ​ ​ 20% until impregnation and embedding are
complete ​ ​ 10 % formalin (12-24 hours)
**All gelatin reagents contain 1 % phenol

ORIENTATION
✔ Arranging the tissue in the mold
✔ Fixing the tissue block on the microtome
✔ Arranging the tissue ribbons on the slide

Types of Embedding Molds


1. Leuckhart’s embedding mold
✔ 2 L-shaped heavy metal arranged flat on a metal
surface
2. Compound E unit
✔ with several interlocking plates making several
compartments
3. Disposable Molds
a. Peel-away (thin plastic Embedding molds)
b. Plastic Ice Trays
c. Paper boats
✔ ADV: cheap, diff sizes, avoids confusion
4. Plastic Embedding Rings & Base Molds

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