I have used many different hier methods over the years and have special considerations because of my altitude. We thought that the pressure cooker would solve all the problems relating to altitude because it is pressurized but it did not. There must be some boiling going on sometime because I often got damaged tissue with the PC. My heat source of choice now includes either a waterbath or steamer, and I use the steamer most often because water boils rapidly above about 92c for me, so I have to keep the waterbath below that. My husband drilled a hole in the lid of the steamer where I insert a thermometer. I use a digital cooking therm., it has a feature that you can set to alarm when it reaches a programmed temp, I put tubs or coplin jars with the buffers in the steamer to warm up while I deparaffinize the slides. I put the temp probe in the solution in the steamer and set it to alarm at 90c. after I put the slides in the buffer in the steamer, I set the alarm again to go off when it again reaches 90c, (it can take up to 10-15 min. to get back to temp after adding the slides) after it reaches 90c with the slides in there I start timing, I steam my slides for 15-20 min after they reach 90+c., they will go up only to about 95c in my lab because of the altitude I assume. When they are done I take them out of the steamer and let them cool on the counter for 20 min or so, then rinse in water and then place in buffer before doing IHC.
Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 [EMAIL PROTECTED] www.ihctech.net <http://www.ihctech.net/> www.ihcrg.org _____ From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Perry, Margaret Sent: Wednesday, October 22, 2008 10:28 AM To: [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu Subject: [IHCRG] HIER We have been using the microwave HIER and have had good results, however when our microwave bits the dust I would like to have a pressure cooker method in place. I feel the pressure cooker is more consistent for all the slides. We are a veterinary diagnostic lab and I would like to have some idea of where to begin. I have looked at different protocols and they often indicate HIER in a pressure cooker but do not give the details. I currently use citrate buffer pH 6. I put the slides in refrigerated buffer and microwave on high for 1 min 45 sec. or until the buffer just starts to boil. I then set the microwave on 10% power for 10 minutes. Afterward the slides are allowed to cool in the microwave for 1 hour. We have a biocare Decloaker Chamber and I would appreciate help with the program I should use. Do you start with cold buffer or should I prewarm it? What temperature should I use? How long should I maintain the temperature? How long should it be before I remove the slides? I also am working with a new protocol that calls for heating in a steamer. Should the temperature of the buffer be warm, cold or room temp when I start? Thank you. Margaret Perry HT (ASCP) IHC Lab Manager Veterinary Science Animal Disease Research and Diagnostic Lab South Dakota State University Box 2175 North Campus Drive Brookings SD 57007 --~--~---------~--~----~------------~-------~--~----~ You received this message because you are subscribed to the Google Groups "ihcrg" group. To post to this group, send email to [EMAIL PROTECTED] To unsubscribe from this group, send email to [EMAIL PROTECTED] For more options, visit this group at http://groups.google.com/group/ihcrg?hl=en -~----------~----~----~----~------~----~------~--~--- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet