Patricia:
I has been always my objective to develop a processing protocol that could be used with all types of tissues and specimens and for all practical urposes I was able to do it. The only tissue I treated differently where those of the CNS that I always run with longer protocols.
My general protocol (for all tissue, and biopsies alike) was:
2 formalin X 30 min. each; 60EthOL + 80EthOL + 95EthOL X 30 min each; 100EthOL s stations (40, 30 and 25 min each); 2 xylenes (25 min each) and 4 paraffin wax (30 min each).
When I stopped using xylene altogether for tissue processing things became really better for all tissues and my general protocol was:
2 formalin (25 min each); 80EthOL 90EthOL and 3 100EthOL (all for 15 min each), EIM (2:3:1) at 45ºC for 1h30min followed by EIM (1:3:2) at 50ºC for 1h 15 min; pure mineral oil at 50ºC for 1 hour and 4 paraffin wax stations (45; 20; 20 and 50 min each).
EIM is a mixture of Ethanol+Isopropanol+Mineral oil in the indicated proportions.
Tissues are processed in a much better way than with xylene and are softer to section, specially the small biopsies, breast, lymph nodes and uterus. I would recommend you to try this procedure.
René J.
Patricia Valente wrote:
I would like to hear about VIP processing times other folks use for prostate biopsies using biopsy sponges.
We currently run an average of 60 cassettes( some days can be 120) all with 2 sponges.- (would prefer to use mesh window cassettes but pathologists prefer the flat and complete sections we (usually) get more easily with the sponges)
We rotate 1 of each reagent type ( 95%,100%, Xylene,wax) on processor after each run - this does seem to help with carryover problem.
Run is 3hrs 45 min. long
Every once in a while we get few nasty hard fragmented cores and other times under dehydration.- These cannot all be blamed on collection or grossing technique ( though we have tried).
Seems like processing is uneven.(PV is on)
Does anyone use longer times with so many sponges?
Has any one tried something to space out cassettes( maybe an empty cassette in-between full ones) to get better contact with solutions?
Any other ideas??
Thanks
Pat Valente
San Antonio TX
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