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FIXATION:        10% buffered neutral formalin

TECHNIQUE:        Paraffin embedded sections cut at 4 μm.

SOLUTIONS:

0.5% Periodic Acid Solution

Periodic acid ---------------------------------------------------------------------        0.5 gm

Distilled water -------------------------------------------------------------------        100.0 ml

Modified Lillie’s Schiff Solution

(see PAS method)

0.3% Sodium Borate

Sodium borate (Na2 B4 O7 10H2O) ----------------------------------------------                0.3 gm

Distilled water -------------------------------------------------------------------        100.0 ml

1. Deparaffinize and hydrate to distilled water.
2. Place in 0.5% periodic acid for 15 minutes.
3. Rinse in four changes of distilled water.
4. Place in modified Lillie’s Schiff solution for 30 minutes.
5. Rinse in four changes distilled water.
6. Place in 0.3% sodium borate for 15 seconds.
7. Rinse in four changes of distilled water.
8. Stain in acidified Lillie-Mayer’s hematoxylin for 45 seconds.
9. Rinse in three changes of distilled water.
10. Blue hematoxylin in 0.3% sodium borate for 15 seconds.
11. Rinse in four changes of distilled water.
12. Dehydrate in graded alcohols.
13. Clear in three or four changes of xylene.
14. Mount with synthetic resin.

PAS positive substances ---------------------------------------------        red

Nuclei ---------------------------------------------------------------        blue

This method gives more intense staining of PAS positive substances.  This is accomplished by oxidizing longer than the usual time in periodic acid and staining longer than usual in modified Lillie’s Schiff reagent.  This intensified staining helps to better visualize the basement membrane of skin biopsies and the bile ducts of transplant liver biopsies.

Garvey, W. et al:  Combined modified periodic acid-Schiff and batch staining method.  J. Histotechnol. 15:117-120, 1992.

Lillie, R.D. and Fullmer, H.W.:  Histopathologic Technic and Practical Histochemistry, 4th Ed., New York, McGraw-Hill, 1976, pp. 206, 307-308.