Re: [Histonet] HIER

From:Larry Woody

I'm wondering if anyone out there has tried something called Uni-Trieve from Innovex Biosciences that claims to be a universal low temperature retrieval solution ie: 75 degrees for 15 minutes. I wouldn't mind trying it on bone sections to see if they stay on better.  Larry A. WoodySeattle, Wa.----- Original Message ----From: Jan Shivers To: "Perry, Margaret" ;; histonet@lists.utsouthwestern.eduSent: Wednesday, October 22, 2008 11:23:05 AMSubject: Re: [Histonet] HIERIn my opinion/experience:The upside of using a microwave for HIER is that you can remove some slides at certain intervals, and keep heating the rest (replacing the removed slides with blanks, so as to keep the same amount of material inside the machine).  My microwave HIER protocols vary in length from 5' up to 20' (in 5' increments, replacing buffer volume if needed), so being able to remove at the right times is essential.  I often have 5-6 different times and buffers going, and could not spend the extra time trying to do HIER separately on each one.  The downside to using a microwave is the verifiability of temperature, if you don't have a fancy laboratory microwave.  I'm currently looking into the various models and price ranges out there for a new one that will record temps, etc.=0ASteam retrieval in a vegetable steamer is great.  However, the downside to steam retrieval is the amount of time needed to heat up the slides/solutions.  Sixty minutes is just too long for my turn-around time. =0AMost days, I think 20 minutes is too long!Pressure cooker HIER works really well, too, but you run into a problem if all your tests for that day don't use the same amount of time in HIER.  Once you break that seal to remove some of the slides that need a short heating time, your temp/pressure are gone.I work in an animal diagnostic lab, and our usual workload is about 150 slides/day, so we need to get the slides in and out fairly rapidly.  Thus, the reason why microwaving works best for me.  Others will have different needs and time allowances in their labs.P.S.  a) I always start with cold buffer prior to heat retrieval.  b)  I always cooldown in HIER buffers for 20'.Jan Shivers=0AUMN VDL----- Original Message ----- From: "Perry, Margaret" To: ; Sent: Wednesday, October 22, 2008 11:28 AMSubject: [Histonet] HIERWe 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 ScienceAnimal Disease Research and Diagnostic LabSouth Dakota State UniversityBox 2175 North Campus DriveBrookings SD 57007_______________________________________________Histonet mailing listHistonet@lists.utsouthwestern.edu mailing listHistonet@lists.utsouthwestern.edu
Histonet mailing list

<< Previous Message | Next Message >>