Rapid hand processing for RUSH biopsies

	Speciemen:  Small biopsies of renal,  cardiac, lung  or hepatic
tissue which have been adequately fixed.  These biopsies should not exceed 2
mm in maximum thickness.

	1.  It is most efficient to collect the biopsy in alcoholic
formalin, thus shortening additional fixation time The specimen is
described, wrapped, stained with eosin  and placed in Unicassettes
(cassettes without metal covers) in alcoholic formalin to which a few drops
of eosin has been added. If the are received fresh. they must remain in
alcoholic formalin for 10 to 15 minutes to ensure adequate fixation.

	2.  Obtain three 250 ml jars with tight fitting lids. Label the
jars: Absolute ethanol,  Acetone I and Acetone II. Place fresh absolute
ethanol into the correctly labeled jar and fresh acetone into the others.
Magnetic stirring bars should be placed into each jar.  A fourth jar or 500
ml beaker should be reserved for molten paraffin.

	3. 	Place the jars labeled "Absolute ethanol" on an unheated
magnetic stirrer - hot plate.  Adjust the stirring action to provide brisk
movement of the fluid. Transfer the cassette(s) from the alcoholic formalin
directly into this jar. Time for 30 seconds.

	4. 	Transfer from the ethanol into the jar labeled Acetone I.
Place on an unheated  magnetic stirrer - hot plate for 3 minutes.

	5.	Transfer from the Acetone I. into the jar labeled Acetone
II.  Place on an unheated  magnetic stirrer - hot plate for 3 minutes.

	6.	While the cassettes are in the last acetone, obtain 200 ml
molten infiltration medium from the appropriate dispenser, and place into
the fourth jar/ beaker.  Maintain the temperature at 60oC. (between 3 and 4
on the hot plate dial but monitor with a thermometer.)

	7.	Transfer the cassettes directly into the paraffin from the
acetone.  Continue agitation with the stirring bar for 6 minutes. Maintain
temperature at 60o C.

		8.  Proceed with the procedure for paraffin embedding.


		Ref: McCue, P.A., Santoianni, R. A.: Expediated Handling of
Transplantation Biopsies. Am J. Surg Path. 1988;12
(2):155-157


Michael M. Fredrickson
Technical Director, Anatomic Pathology
Massachusetts General Hospital
Boston, MA 02114