Re: [Histonet] Water under sections
I use charged slides all the time. The best way to clear the water is to cut the slides and air dry them overnight. The next day I follow the normal protocol, heat in 60 degree over and then on to the rest of the protocol. They don't really need the 56 degree oven all night long. Respectfully, Colleen Forster On Mon, Jan 25, 2021 at 11:26 AM Tyrone Genade via Histonet < histonet@lists.utsouthwestern.edu> wrote: > Hello, > > > On Saturday, January 23, 2021 1:00 PM, < > histonet-requ...@lists.utsouthwestern.edu> wrote: > > > In my lab, water under the sections is unique to charged slides. > > And you > > are correct, if there is water under the section when the slides > > are heated > > for antigen retrieval, the boiling (or at least very hot) water > > will damage > > or entirely destroy the section. > > I had this very problem and the simple solution was to bake the slides > overnight at 56oC (melting temp of the wax I was working with). I would > still heat on a hot plate before dewaxing. Baking at 45oC also seemed to > work. Whether this would work for you would depend on the sensitivity of > your epitopes to prolonged heat. > > Good luck... every now and then a section would still detach during > antigen retrieval. > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Colleen Forster HT(ASCP)QIHC BLS Histology and IHC Laboratory Jackson Hall, Room 2-155 612-626-1930 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Water under sections
Hello, On Saturday, January 23, 2021 1:00 PM, wrote: > In my lab, water under the sections is unique to charged slides. > And you > are correct, if there is water under the section when the slides > are heated > for antigen retrieval, the boiling (or at least very hot) water > will damage > or entirely destroy the section. I had this very problem and the simple solution was to bake the slides overnight at 56oC (melting temp of the wax I was working with). I would still heat on a hot plate before dewaxing. Baking at 45oC also seemed to work. Whether this would work for you would depend on the sensitivity of your epitopes to prolonged heat. Good luck... every now and then a section would still detach during antigen retrieval. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Water under sections
This is all good advise. I always picked up section and tapped the slide on counter and then dried standing up. This is really a problem if you are at all above 5K feet in altitude where water boils violently at 92-93dc, which will destroy your tissue section. I have written a couple of papers about on this. From: Greg Dobbin Sent: Friday, January 22, 2021 11:45 AM To: b-freder...@northwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Water under sections Hi Bernice, In my lab, water under the sections is unique to charged slides. And you are correct, if there is water under the section when the slides are heated for antigen retrieval, the boiling (or at least very hot) water will damage or entirely destroy the section. We allow the charged slides to drain (upright) for a few minutes and then carefully make a hole in the edge of the wax and use a Kimwipe or paper towel to carefully wick the excess water out as much as we can without touching the tissue section. Then we "flick" or shake sharply to remove any residual water that may remain before we bake them (FYI, we choose not to bake on the stainer). If after baking 30 mins at 60C, we notice water (maybe someone was not as diligent earlier?), we wick away the excess and bake for another 15 mins to ensure good adhesion of the section to the slide before proceeding to the immunostainer. Hope this helps. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Water under sections
Hi Bernice, In my lab, water under the sections is unique to charged slides. And you are correct, if there is water under the section when the slides are heated for antigen retrieval, the boiling (or at least very hot) water will damage or entirely destroy the section. We allow the charged slides to drain (upright) for a few minutes and then carefully make a hole in the edge of the wax and use a Kimwipe or paper towel to carefully wick the excess water out as much as we can without touching the tissue section. Then we "flick" or shake sharply to remove any residual water that may remain before we bake them (FYI, we choose not to bake on the stainer). If after baking 30 mins at 60C, we notice water (maybe someone was not as diligent earlier?), we wick away the excess and bake for another 15 mins to ensure good adhesion of the section to the slide before proceeding to the immunostainer. Hope this helps. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
