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Staining and Collodionization
This document discusses procedures for staining tissue sections mounted on paraffin or celloidin slides. It outlines the general staining process which involves deparaffinization or clearing in xylene or alcohol, hydration in graded alcohols, and staining. Special considerations are given for staining celloidin sections which are soluble in alcohol. The process of collodionization is described as a way to reinforce sections mounted on paraffin ribbons that contain air bubbles, tears or inadequate infiltration.
Staining and Collodionization
- 1.
- 2. General Staining Procedure MOUNT Xylene(1) Xylene DEPARAFFINIZATION CLEARING Xylene(1I) Xylene Absolute Alcohol (I) Absolute Alcohol (I) Absolute Alcohol (II) Absolute Alcohol (II) SECTIONS TO DEHYDRATION ALCOHOL 90% Alcohol 90% Alcohol 70% Alcohol 70% Alcohol Water STAIN
- 3. Staining of ParaffinSections
- 4. Staining of ParaffinSections MOUNT Xylene(1) Xylene DEPARAFFINIZATION CLEARING Xylene(1I) Xylene Absolute Alcohol (I) Absolute Alcohol (I) Absolute Alcohol (II) Absolute Alcohol (II) SECTIONS TO DEHYDRATION ALCOHOL 90% Alcohol 90% Alcohol 70% Alcohol 70% Alcohol Water STAIN
- 5. Staining of CelloidinSections
- 6. Staining of CelloidinSections Cellulose Nitrate Soluble in alcohol, treatment avoided PRECAUTIONS ◦ Stains on skin ◦ Dirty or oily slide ◦ Faulty staining solution ◦ Too much adhesive
- 7. Collodionization of Sections Definition If paraffin ribbons ◦ Contain air bubbles ◦ Torn ◦ Inadequately infiltrated Purpose Procedure
Editor's Notes
- #3 After section cutting, and mounting on the slide , drained , dried to ensure firm attachment of section to slide
- #5 Xylene, a solvent, 2x at 1-2 mins eachXylene not miscible with alcohol (DECREASING CONC)alcohol then replaced with water ,except if use alcoholic stainsincreasing conc for dehydration because most mountants are miscible with alcoholsecond xylene for refractive index, may stay there for an indefinite period until section is mounted on slideFloat off :: dirty or greasy slides,wet
- #7 RemediesTopical application of 0.5% acid alcohol, then rinse with tap waterAlbumin fixative too old, to avoid, adhesives should be prepared in not too great amounts, last 2-3 monthsSection does not stain, prevent by sufficient ripened Hematoxylin, Impurities, improper washing off of paraffin, fixative, decalcifying solutionSections not clear under the microscope, xylol used to dissolve