Rapid tissue processing system
A method for processing a biological tissue specimen using either manual or automated processing. The tissue specimen is sequentially contacted with components of a reagent system in the presence of increased temperature and/or agitation and in the absence of exogenous microwave radiation to facilitate processing in less than one working day.
1. A method for histologically processing a solid biological tissue specimen for histopathological evaluation, the method comprising,
(a) contacting a biological tissue specimen equal or <5.0 mm thick with a fixative solution comprising (i) formalin buffered to a substantially neutral pH, and (ii) at least about 95% (v/v) ethanol, at about 55° C. to about 57° C.; then,
(b) contacting said tissue specimen with a dehydration solution comprising about 90% (v/v) isopropanol and about 9% (v/v) methanol, at about 55° C. to about 57° C.; and then,
(c) contacting said tissue specimen with a clearing solution,
wherein the processing of said biological tissue specimen equal or <5.0 mm thick in the presence of centrifugal agitation at a temperature not exceeding 57° C. without exposing the tissue specimen to microwave radiation is in less than about five hours.
2. The method of claim 1 wherein the tissue specimen is contacted with a holding solution for at least 20 minutes prior to contact with the fixative solution.
3. The method of claim 2 wherein the tissue specimen is contacted with a holding solution prior to the fixative solution at about 20° to about 22°.
4. The method of claim 1 wherein the tissue specimen is contacted with an infiltrating solution in the presence of centrifugal agitation after the clearing solution at about 62° C. to about 64° C.
5. The method of claim 2 wherein the holding solution comprises a formaldehyde-based solution.
6. The method of claim 4 wherein the infiltrating solution comprises paraffin.
7. The method of claim 2 wherein the holding solution further comprises at least one reagent that facilitates penetration of the fixation solution into the tissue specimen.
8. The method of claim 2 wherein the holding solution further comprises at least one of a non-ionic detergent or an aprotic solvent.
9. The method of claim 8 wherein the non-ionic detergent is selected from the group consisting of Triton-X 100, Tween 20, Nonidet-P40, and combinations thereof.
10. The method of claim 8 wherein the aprotic solvent is dimethyl sulfoxide (DMSO).
11. The method of claim 1 wherein the fixative solution further comprises at least one of polyethylene glycol or DMSO.
12. The method of claim 1 wherein the dehydration solution further comprises about 0.5% (v/v) formaldehyde.
13. The method of claim 1 wherein the dehydration solution further comprises about 0.1% (v/v) PEG and about 0.4% (v/v) DMSO.