drying and coverslipping machines....RE: [Histonet] RE: air drying special stain slides rather than
If you dry the slides do you coverslip on a coverslipping machine - in which you have to put into xylene to run? Tim Morken -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lori Harris Sent: Tuesday, September 11, 2012 10:23 AM To: E. Wayne Johnson; Rene J Buesa Cc: 'histonet@lists.utsouthwestern.edu'; Mayer, Toysha N Subject: RE: [Histonet] RE: air drying special stain slides rather than We have been using the oven dry method for special stains for about four years now and it works wonderfully. Lori A. Harris, HT (ASCP) Histology Section Lead GSRMC Pathology Lab 3600 NW Samaritan Drive Corvallis, OR 97330 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of E. Wayne Johnson Sent: Tuesday, September 11, 2012 10:15 AM To: Rene J Buesa Cc: 'histonet@lists.utsouthwestern.edu'; Mayer, Toysha N Subject: Re: [Histonet] RE: air drying special stain slides rather than I am convinced to give it a try because I also have trouble will the loss of some stains in dehydration. I was concerned that the slides would not clear well after oven dehydration. I will see how it works for me. I can see clearly how going from counterstain to oven will save much hassle with xylene and alcohols as well as not washing out some special stains. I have tried some of the isopropyl alcohol and acetone dehydration called for in some of the stain procedures and it would be great if the slides could just be popped into the oven. What mounting medium are you using? Does it matter? I am a bit worried about penetration of the mountant into the tissue section if there is no xylene in the tissue. Will neutral balsam still work ok? Rene: if you have a link to the paper you talked about on eliminating xylene, I am interested. Xylene is becoming more and more of an issue and a pain for us. EWJohnson Enruikang Ag Tech Beijing. On 9/12/2012 12:01 AM, Rene J Buesa wrote: Toysha: Perhaps you have not oven dried stained slides before, and that explains some of your comments, like: 1- if the stained slides are completely dried, the miscibility you point out is not an issues, because there is nothing to mix with; 2- if you dehydrate → clear the stained sections that will take about 15 minutes per group of up to 25 slides, or even more depending on the protocol used in your automated stainer, but if your group of slides in their rack are placed in an oven at 60ºC for 5 minutes it will just that, 5 minutes reducing the usual TAT for each staining procedure; 3- any oven can accommodate more than 100 stained slides in their racks and the TAT is shortened by oven drying, no matter how many slides you are working with; 4- I really do not know where you can find that extreme heat can affect the tissue sections. All tissue sections are fixed → processed → dried (usually at the same 60ºC before staining) → stained and an additional step at 60ºC to dry before cover-slipping is just that, an additional step at 60ºC 5- The so called Lean technologies do not refer to staining only, they have to do with the whole work-flow and an additional drying step at 60ºC cannot affect in a negative way to the work-flow 6- after staining you will oven dry the sections. I think you should try the method instead. René J. From: Mayer,Toysha Ntnma...@mdanderson.org To: 'histonet@lists.utsouthwestern.edu'histo...@lists.utsouthwestern.ed u Sent: Tuesday, September 11, 2012 11:41 AM Subject: [Histonet] RE: air drying special stain slides rather than Ooh, great question for my students next semester. Your answer is the counterstain, some counterstains may require dehydration after rinsing, or some may not. Adjusting the times of the counterstain is not the issue as much as the solvent of the counterstain. Rene, while I do acknowledge that the xylene may/will cause hazards, we must think of the miscibility of the clearant and the dehydrant, as well as the amount of time involved. The amount of time involved to blot and air dry the slides will affect the TAT for the specimen. 5 min may be ok if you have a small amount of slides, but with a larger number of slides, it will be considerably more than 5. Also Lean methodologies would not apply in that case. With automation, the extreme heat involved with a stain dryer may affect the tissue on the slide. There are some stains that can be blotted, cleared and coverslipped, but using the alcohol to remove excess water and counter stain is better in my opinion. Toysha N. Mayer, MBA, HT (ASCP) Instructor, Education Coordinator Program in Histotechnology School of Health Professions MD Anderson Cancer Center (713) 563-3481 tnma...@mdanderson.org Message: 16 Date: Tue, 11 Sep 2012 10:32:08 -0400 From: Diana
Re: drying and coverslipping machines....RE: [Histonet] RE: air drying special stain slides rather than
I have used Sakura film and Leica glass coverslippers (slides directly from the oven).. René J. From: Morken, Timothy timothy.mor...@ucsfmedctr.org To: Lori Harris lhar...@samhealth.org; E. Wayne Johnson e...@pigsqq.org; Rene J Buesa rjbu...@yahoo.com Cc: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu; Mayer, Toysha N tnma...@mdanderson.org Sent: Tuesday, September 11, 2012 1:30 PM Subject: drying and coverslipping machinesRE: [Histonet] RE: air drying special stain slides rather than If you dry the slides do you coverslip on a coverslipping machine - in which you have to put into xylene to run? Tim Morken -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lori Harris Sent: Tuesday, September 11, 2012 10:23 AM To: E. Wayne Johnson; Rene J Buesa Cc: 'histonet@lists.utsouthwestern.edu'; Mayer, Toysha N Subject: RE: [Histonet] RE: air drying special stain slides rather than We have been using the oven dry method for special stains for about four years now and it works wonderfully. Lori A. Harris, HT (ASCP) Histology Section Lead GSRMC Pathology Lab 3600 NW Samaritan Drive Corvallis, OR 97330 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of E. Wayne Johnson Sent: Tuesday, September 11, 2012 10:15 AM To: Rene J Buesa Cc: 'histonet@lists.utsouthwestern.edu'; Mayer, Toysha N Subject: Re: [Histonet] RE: air drying special stain slides rather than I am convinced to give it a try because I also have trouble will the loss of some stains in dehydration. I was concerned that the slides would not clear well after oven dehydration. I will see how it works for me. I can see clearly how going from counterstain to oven will save much hassle with xylene and alcohols as well as not washing out some special stains. I have tried some of the isopropyl alcohol and acetone dehydration called for in some of the stain procedures and it would be great if the slides could just be popped into the oven. What mounting medium are you using? Does it matter? I am a bit worried about penetration of the mountant into the tissue section if there is no xylene in the tissue. Will neutral balsam still work ok? Rene: if you have a link to the paper you talked about on eliminating xylene, I am interested. Xylene is becoming more and more of an issue and a pain for us. EWJohnson Enruikang Ag Tech Beijing. On 9/12/2012 12:01 AM, Rene J Buesa wrote: Toysha: Perhaps you have not oven dried stained slides before, and that explains some of your comments, like: 1- if the stained slides are completely dried, the miscibility you point out is not an issues, because there is nothing to mix with; 2- if you dehydrate → clear the stained sections that will take about 15 minutes per group of up to 25 slides, or even more depending on the protocol used in your automated stainer, but if your group of slides in their rack are placed in an oven at 60ºC for 5 minutes it will just that, 5 minutes reducing the usual TAT for each staining procedure; 3- any oven can accommodate more than 100 stained slides in their racks and the TAT is shortened by oven drying, no matter how many slides you are working with; 4- I really do not know where you can find that extreme heat can affect the tissue sections. All tissue sections are fixed → processed → dried (usually at the same 60ºC before staining) → stained and an additional step at 60ºC to dry before cover-slipping is just that, an additional step at 60ºC 5- The so called Lean technologies do not refer to staining only, they have to do with the whole work-flow and an additional drying step at 60ºC cannot affect in a negative way to the work-flow 6- after staining you will oven dry the sections. I think you should try the method instead. René J. From: Mayer,Toysha Ntnma...@mdanderson.org To: 'histonet@lists.utsouthwestern.edu'histo...@lists.utsouthwestern.ed u Sent: Tuesday, September 11, 2012 11:41 AM Subject: [Histonet] RE: air drying special stain slides rather than Ooh, great question for my students next semester. Your answer is the counterstain, some counterstains may require dehydration after rinsing, or some may not. Adjusting the times of the counterstain is not the issue as much as the solvent of the counterstain. Rene, while I do acknowledge that the xylene may/will cause hazards, we must think of the miscibility of the clearant and the dehydrant, as well as the amount of time involved. The amount of time involved to blot and air dry the slides will affect the TAT for the specimen. 5 min may be ok if you have a small amount of slides, but with a larger number of slides, it will be considerably more than 5. Also Lean methodologies would
