RE: [Histonet] Histogel
Thank you, John! For some strange reason, the URL in your message got garbled/multiplied, but I just took this part: http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf and that worked. The enrobing I do is to trap debris on the surface of a coral skeleton at the interface between apparently healthy tissue and bare skeleton in studying the tissue loss diseases of the corals. After solidifying the agarose, I decal the sample in neutral pH 10% EDTA, then cut (breadloaf-style) the agarose-enrobed tissue block into ~2-mm slices and put each in a cassette for processing. Perhaps my problem is the processing time needs to be increased to be sure to remove all the water from the agarose and infiltrate it completely. I don't know what the instructions mean by non-porous filter paper, either, has anyone figured this out? Esther From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of John Kiernan jkier...@uwo.ca Sent: Tuesday, January 21, 2014 8:54 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histogel The document at http://lists.utsouthwestern.edu/mailman/listinfo/histonet(http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf;http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf gives a thorough description of Histogel, and even says what it is - hydroxyethyl agarose. In the detailed instructions for various uses, the only confusing thing is the requirement for non-porous filter paper! John Kiernan Anatomy, UWO London, Canada = = = On 20/01/14, Elizabeth Chlipala l...@premierlab.com wrote: Esther I agree with Dusko, I fix before I put in histogel and again after the sample is placed in histogel, we have no formalin on our tissue processor, we start in 50% alcohol. I also process on a longer processing cycle, 1 hour per station and similar to Dusko's - denatured ethanol, xylene and paraplast and paraplast extra to embed. I've never had a problem (such as overprocessed tissue) with the histogel or the sample embedded in the histogel with the longer processing cycle. Most of the samples we process are cell blocks or tissue fragments such as micronized tissue constructs, which are like powder when we receive them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dusko trajkovic Sent: Monday, January 20, 2014 12:47 PM To: Esther C Peters; jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Esther, I mainly process cells, which have been spun down into a small pellet. Also mouse DRG's and other very small tissues. I would consider this delicate, so do not be afraid to use a longer processing program. Histogel/Agurose is what needs longer dehydrating steps. We do not use any substitute reagents, so in that aspect I cannot tell you how they will affect the processing. Our lab uses ethanol, xylene, and Paraplast paraffin. Try a test run and let me know if you were able to get successful results. Have a good Monday! Dusko From: Esther C Peters epete...@gmu.edu To: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 11:15 AM Subject: RE: [Histonet] Histogel Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu
RE: [Histonet] Histogel
The document at http://lists.utsouthwestern.edu/mailman/listinfo/histonet(http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf;http://www.thermo.com/eThermo/CMA/PDFs/Various/File_9759.pdf gives a thorough description of Histogel, and even says what it is - hydroxyethyl agarose. In the detailed instructions for various uses, the only confusing thing is the requirement for non-porous filter paper! John Kiernan Anatomy, UWO London, Canada = = = On 20/01/14, Elizabeth Chlipala l...@premierlab.com wrote: Esther I agree with Dusko, I fix before I put in histogel and again after the sample is placed in histogel, we have no formalin on our tissue processor, we start in 50% alcohol. I also process on a longer processing cycle, 1 hour per station and similar to Dusko's - denatured ethanol, xylene and paraplast and paraplast extra to embed. I've never had a problem (such as overprocessed tissue) with the histogel or the sample embedded in the histogel with the longer processing cycle. Most of the samples we process are cell blocks or tissue fragments such as micronized tissue constructs, which are like powder when we receive them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dusko trajkovic Sent: Monday, January 20, 2014 12:47 PM To: Esther C Peters; jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Esther, I mainly process cells, which have been spun down into a small pellet. Also mouse DRG's and other very small tissues. I would consider this delicate, so do not be afraid to use a longer processing program. Histogel/Agurose is what needs longer dehydrating steps. We do not use any substitute reagents, so in that aspect I cannot tell you how they will affect the processing. Our lab uses ethanol, xylene, and Paraplast paraffin. Try a test run and let me know if you were able to get successful results. Have a good Monday! Dusko From: Esther C Peters epete...@gmu.edu To: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 11:15 AM Subject: RE: [Histonet] Histogel Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Jennifer, You might have seen one of my posts from 2-3 years ago. I had the exact same problems you described. Could not get anyone to come up with a solution. I ran various programs on our VIP and finally came up with a solution. Fix your specimens as you normally would do. Drain of the fixative add your histogel (dissolved in hot water, which you have been doing), fill the mold with the histogel. Let solidify on ice or 4C in fridge. Place the solid histogel in a cassette and process on a 12 hour program. Since I have instituted this procedure, have not had one bad block to date. Longer processing is the answer, and nothing else. Good Luck. Dusko Trajkovic Pfizer Inc. La Jolla 858-638-6202 From: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com To: histonet@lists.utsouthwestern.edu Sent: Monday, January 20, 2014 7:56 AM Subject: [Histonet] Histogel Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had
[Histonet] Histogel
Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had problems with it - some samples would come out great and some would shrivel up or even dissolve. These posts on the Histogel were from a few years ago and was hoping, and praying that someone out there may have solved this issue and have a little info you could share with me on this subject. Did anyone out there ever figure out how to get consistent results? I have spoken with RA Scientific and they have no additional insights. Here is the background: I have used Histogel for about 4 years now. In September of last year, we started seeing the shriveling Histogel samples. Like others who posted, it was random. I could embed two serial pieces of nerve, from the same mouse, into two blocks and one would shrivel and one would look great. So I have tried many things, always in multiples of 3 or more per condition per run... Fixing in formalin only, embedding in Histogel and storing in PBS until processing Fixing in formalin only, embedding in Histogel and storing in 40% reagent alcohol until processing (the first step of our processor is 40%) Fixing in formalin only, embedding in Histogel and storing in formalin until processing Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing in 40% or formalin until processing For all of these conditions, I have tried using a small cycle (30 min/bath) and a biopsy cycle (15 minutes/bath). Once processed, there was no rhyme or reason to the results. Some blocks looked great; others within the same group looked shriveled. Sometimes the blocks were white, sometimes they were clear. Next, I thought it was my pre-processing - so I heated the Histogel in a water bath, rather than microwaving. That way all of the samples were embedded with the Histogel at the same temperature - about 55 degrees. Again, no rhyme or reason, some looked good, some looked bad. Lastly I thought that maybe I was carrying over too much liquid from my sample to the Histogel so I tried the following: I made Histogel only blocks or added 3-4 drops of 40% alcohol or formalin to the liquid Histogel before a tissue/cell free block was made. Yep, you guessed it - no luck. Some looked good, some looked shriveled. So here I have this great tool to embed tiny samples, but I am afraid to use it because I don't know if it will work or shrivel! Can anyone out there help me? Thanks, Jenn Jennifer Johnson Staff Scientist Genzyme, a Sanofi Company Department of Pathology 5 Mountain Road Framingham, MA 01701-9322 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Histogel
Jennifer, You might have seen one of my posts from 2-3 years ago. I had the exact same problems you described. Could not get anyone to come up with a solution. I ran various programs on our VIP and finally came up with a solution. Fix your specimens as you normally would do. Drain of the fixative add your histogel (dissolved in hot water, which you have been doing), fill the mold with the histogel. Let solidify on ice or 4C in fridge. Place the solid histogel in a cassette and process on a 12 hour program. Since I have instituted this procedure, have not had one bad block to date. Longer processing is the answer, and nothing else. Good Luck. Dusko Trajkovic Pfizer Inc. La Jolla 858-638-6202 From: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com To: histonet@lists.utsouthwestern.edu Sent: Monday, January 20, 2014 7:56 AM Subject: [Histonet] Histogel Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had problems with it - some samples would come out great and some would shrivel up or even dissolve. These posts on the Histogel were from a few years ago and was hoping, and praying that someone out there may have solved this issue and have a little info you could share with me on this subject. Did anyone out there ever figure out how to get consistent results? I have spoken with RA Scientific and they have no additional insights. Here is the background: I have used Histogel for about 4 years now. In September of last year, we started seeing the shriveling Histogel samples. Like others who posted, it was random. I could embed two serial pieces of nerve, from the same mouse, into two blocks and one would shrivel and one would look great. So I have tried many things, always in multiples of 3 or more per condition per run... Fixing in formalin only, embedding in Histogel and storing in PBS until processing Fixing in formalin only, embedding in Histogel and storing in 40% reagent alcohol until processing (the first step of our processor is 40%) Fixing in formalin only, embedding in Histogel and storing in formalin until processing Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing in 40% or formalin until processing For all of these conditions, I have tried using a small cycle (30 min/bath) and a biopsy cycle (15 minutes/bath). Once processed, there was no rhyme or reason to the results. Some blocks looked great; others within the same group looked shriveled. Sometimes the blocks were white, sometimes they were clear. Next, I thought it was my pre-processing - so I heated the Histogel in a water bath, rather than microwaving. That way all of the samples were embedded with the Histogel at the same temperature - about 55 degrees. Again, no rhyme or reason, some looked good, some looked bad. Lastly I thought that maybe I was carrying over too much liquid from my sample to the Histogel so I tried the following: I made Histogel only blocks or added 3-4 drops of 40% alcohol or formalin to the liquid Histogel before a tissue/cell free block was made. Yep, you guessed it - no luck. Some looked good, some looked shriveled. So here I have this great tool to embed tiny samples, but I am afraid to use it because I don't know if it will work or shrivel! Can anyone out there help me? Thanks, Jenn Jennifer Johnson Staff Scientist Genzyme, a Sanofi Company Department of Pathology 5 Mountain Road Framingham, MA 01701-9322 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Histogel
Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Jennifer, You might have seen one of my posts from 2-3 years ago. I had the exact same problems you described. Could not get anyone to come up with a solution. I ran various programs on our VIP and finally came up with a solution. Fix your specimens as you normally would do. Drain of the fixative add your histogel (dissolved in hot water, which you have been doing), fill the mold with the histogel. Let solidify on ice or 4C in fridge. Place the solid histogel in a cassette and process on a 12 hour program. Since I have instituted this procedure, have not had one bad block to date. Longer processing is the answer, and nothing else. Good Luck. Dusko Trajkovic Pfizer Inc. La Jolla 858-638-6202 From: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com To: histonet@lists.utsouthwestern.edu Sent: Monday, January 20, 2014 7:56 AM Subject: [Histonet] Histogel Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had problems with it - some samples would come out great and some would shrivel up or even dissolve. These posts on the Histogel were from a few years ago and was hoping, and praying that someone out there may have solved this issue and have a little info you could share with me on this subject. Did anyone out there ever figure out how to get consistent results? I have spoken with RA Scientific and they have no additional insights. Here is the background: I have used Histogel for about 4 years now. In September of last year, we started seeing the shriveling Histogel samples. Like others who posted, it was random. I could embed two serial pieces of nerve, from the same mouse, into two blocks and one would shrivel and one would look great. So I have tried many things, always in multiples of 3 or more per condition per run... Fixing in formalin only, embedding in Histogel and storing in PBS until processing Fixing in formalin only, embedding in Histogel and storing in 40% reagent alcohol until processing (the first step of our processor is 40%) Fixing in formalin only, embedding in Histogel and storing in formalin until processing Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing in 40% or formalin until processing For all of these conditions, I have tried using a small cycle (30 min/bath) and a biopsy cycle (15 minutes/bath). Once processed, there was no rhyme or reason to the results. Some blocks looked great; others within the same group looked shriveled. Sometimes the blocks were white, sometimes they were clear. Next, I thought it was my pre-processing - so I heated the Histogel in a water bath, rather than microwaving. That way all of the samples were embedded with the Histogel at the same temperature - about 55 degrees. Again, no rhyme or reason, some looked good, some looked bad. Lastly I thought that maybe I was carrying over too much liquid from my sample to the Histogel so I tried the following: I made Histogel only blocks or added 3-4 drops of 40% alcohol or formalin to the liquid Histogel before a tissue/cell free block was made. Yep, you guessed it - no luck. Some looked good, some looked shriveled. So here I have this great tool to embed tiny samples, but I am afraid to use it because I don't know if it will work or shrivel! Can anyone out there help me? Thanks, Jenn Jennifer Johnson Staff Scientist Genzyme, a Sanofi Company Department of Pathology 5 Mountain Road Framingham, MA 01701-9322 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo
Re: [Histonet] Histogel
Esther, I mainly process cells, which have been spun down into a small pellet. Also mouse DRG's and other very small tissues. I would consider this delicate, so do not be afraid to use a longer processing program. Histogel/Agurose is what needs longer dehydrating steps. We do not use any substitute reagents, so in that aspect I cannot tell you how they will affect the processing. Our lab uses ethanol, xylene, and Paraplast paraffin. Try a test run and let me know if you were able to get successful results. Have a good Monday! Dusko From: Esther C Peters epete...@gmu.edu To: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 11:15 AM Subject: RE: [Histonet] Histogel Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Jennifer, You might have seen one of my posts from 2-3 years ago. I had the exact same problems you described. Could not get anyone to come up with a solution. I ran various programs on our VIP and finally came up with a solution. Fix your specimens as you normally would do. Drain of the fixative add your histogel (dissolved in hot water, which you have been doing), fill the mold with the histogel. Let solidify on ice or 4C in fridge. Place the solid histogel in a cassette and process on a 12 hour program. Since I have instituted this procedure, have not had one bad block to date. Longer processing is the answer, and nothing else. Good Luck. Dusko Trajkovic Pfizer Inc. La Jolla 858-638-6202 From: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com To: histonet@lists.utsouthwestern.edu Sent: Monday, January 20, 2014 7:56 AM Subject: [Histonet] Histogel Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had problems with it - some samples would come out great and some would shrivel up or even dissolve. These posts on the Histogel were from a few years ago and was hoping, and praying that someone out there may have solved this issue and have a little info you could share with me on this subject. Did anyone out there ever figure out how to get consistent results? I have spoken with RA Scientific and they have no additional insights. Here is the background: I have used Histogel for about 4 years now. In September of last year, we started seeing the shriveling Histogel samples. Like others who posted, it was random. I could embed two serial pieces of nerve, from the same mouse, into two blocks and one would shrivel and one would look great. So I have tried many things, always in multiples of 3 or more per condition per run... Fixing in formalin only, embedding in Histogel and storing in PBS until processing Fixing in formalin only, embedding in Histogel and storing in 40% reagent alcohol until processing (the first step of our processor is 40%) Fixing in formalin only, embedding in Histogel and storing in formalin until processing Fixing in formalin, rinsing in 40% alcohol, embedding in Histogel, and storing in 40% or formalin until processing For all of these conditions, I have tried using a small cycle (30 min/bath) and a biopsy cycle (15 minutes/bath). Once processed, there was no rhyme or reason to the results. Some blocks looked great; others within the same group looked shriveled. Sometimes the blocks were white, sometimes they were clear. Next, I thought it was my pre-processing - so I heated the Histogel in a water bath, rather than microwaving. That way all of the samples were embedded with the Histogel at the same temperature - about 55 degrees. Again, no rhyme or reason, some looked good, some looked bad. Lastly I thought that maybe I was carrying over too much liquid from my sample
RE: [Histonet] Histogel
Esther I agree with Dusko, I fix before I put in histogel and again after the sample is placed in histogel, we have no formalin on our tissue processor, we start in 50% alcohol. I also process on a longer processing cycle, 1 hour per station and similar to Dusko's - denatured ethanol, xylene and paraplast and paraplast extra to embed. I've never had a problem (such as overprocessed tissue) with the histogel or the sample embedded in the histogel with the longer processing cycle. Most of the samples we process are cell blocks or tissue fragments such as micronized tissue constructs, which are like powder when we receive them. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dusko trajkovic Sent: Monday, January 20, 2014 12:47 PM To: Esther C Peters; jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Esther, I mainly process cells, which have been spun down into a small pellet. Also mouse DRG's and other very small tissues. I would consider this delicate, so do not be afraid to use a longer processing program. Histogel/Agurose is what needs longer dehydrating steps. We do not use any substitute reagents, so in that aspect I cannot tell you how they will affect the processing. Our lab uses ethanol, xylene, and Paraplast paraffin. Try a test run and let me know if you were able to get successful results. Have a good Monday! Dusko From: Esther C Peters epete...@gmu.edu To: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu; dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 11:15 AM Subject: RE: [Histonet] Histogel Thank you, Dusko! I have had the same problem with 1.5% agarose, and I tried starting the dehydration with 30% to 50% to 70% ethanol, and using different xylene substitutes. It appears that the variable whitening and shrinking happens after 100% reagent alcohol and in the xylene substitute (now using Richard-Allan Clear-Rite3). I've wondered if slow infiltration was the issue. I guess we'll try this longer processing, but I also work with delicate tissues that normally would be a short run (15 min in each reagent). Are your tissues thin/delicate/biopsy or cell preps or organ samples? No effect on them? Esther Esther C. Peters, Ph.D. Assistant Professor Environmental Science Policy George Mason University 4400 University Drive, MS 5F2 Fairfax, VA 22030- From: histonet-boun...@lists.utsouthwestern.edu histonet-boun...@lists.utsouthwestern.edu on behalf of dusko trajkovic dunat...@sbcglobal.net Sent: Monday, January 20, 2014 1:58 PM To: jennifer.arcand-john...@genzyme.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histogel Jennifer, You might have seen one of my posts from 2-3 years ago. I had the exact same problems you described. Could not get anyone to come up with a solution. I ran various programs on our VIP and finally came up with a solution. Fix your specimens as you normally would do. Drain of the fixative add your histogel (dissolved in hot water, which you have been doing), fill the mold with the histogel. Let solidify on ice or 4C in fridge. Place the solid histogel in a cassette and process on a 12 hour program. Since I have instituted this procedure, have not had one bad block to date. Longer processing is the answer, and nothing else. Good Luck. Dusko Trajkovic Pfizer Inc. La Jolla 858-638-6202 From: jennifer.arcand-john...@genzyme.com jennifer.arcand-john...@genzyme.com To: histonet@lists.utsouthwestern.edu Sent: Monday, January 20, 2014 7:56 AM Subject: [Histonet] Histogel Dear Histonetters, I have been reading up on the archives for info on Histogel. Previous posts discuss how they had problems with it - some samples would come out great and some would shrivel up or even dissolve. These posts on the Histogel were from a few years ago and was hoping, and praying that someone out there may have solved this issue and have a little info you could share with me on this subject. Did anyone out there ever figure out how to get consistent results? I have spoken with RA Scientific and they have no additional insights. Here is the background: I have used Histogel for about 4 years now. In September of last year, we started seeing the shriveling Histogel samples. Like others who posted, it was random. I could embed two serial pieces of nerve, from the same mouse
[Histonet] Histogel Protocol FROZEN Histonet Digest, Vol 91, Issue 34
Katy (or anyone else who has used Histogel for FROZEN samples), I am using Histogel mixed with OCT to create a frozen cell pellet. I was successful with doing a single pellet but now I am being asked to do multiple and have found hit some challenges. I have never done a TMA but that sounds like a great option. We are a research lab but using the Cell Pellet for a normalizing control of IF intensity (for computational analysis normalization for lung tissue) and one of the largest challenges is retaining cell density while still trying to cryoprotect and freeze well with the OCT surrounding it. Has anyone worked out a method for frozen cell pellets to embedded with multiple cell lines? Most of what I have found has been from FFPE, any suggestions would be appreciated. Thank you! Kathy Kathy M. Bonness, PhD. 251-533-2661 kathy.bonn...@utsouthwestern.edu http://www.linkedin.com/pub/kathy-bonness/6/1a1/931 UTSW Dallas, TX Green Center for Computational Systems Biology Department of Pharmacology Altschuler/Wu Lab (ND9.214) From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of histonet-requ...@lists.utsouthwestern.edu [histonet-requ...@lists.utsouthwestern.edu] Sent: Saturday, June 25, 2011 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 91, Issue 34 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than Re: Contents of Histonet digest... Today's Topics: 1. RE: HistoGel (Milne, Katy) 2. Histogel Problem (Amos Brooks) 3. RE: Histogel Problem (Delossantos_Roseann) 4. bluing (Webb, Dorothy L) 5. Re: bluing (Rene J Buesa) 6. RE: bluing (Harrison, Sandra C.) 7. blades (Webb, Dorothy L) 8. Re: blades (Sean McBride) 9. Re: blades (Esther C Peters) 10. Re: blades (Victoria Baker) 11. Re: blades (Jennifer MacDonald) 12. HT Position - Irvine, CA (Eric Velazquez) 13. Re: HT Position - Irvine, CA (Lee Peggy Wenk) 14. Contents of Histonet digest (Aurea Marquez) 15. Re: bluing (Lee Peggy Wenk) 16. Leica Bond for IHCs (Sheila Adey) 17. RE: Bluing (gayle callis) -- Message: 1 Date: Fri, 24 Jun 2011 10:27:09 -0700 From: Milne, Katy kmi...@bccancer.bc.ca Subject: [Histonet] RE: HistoGel To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu,'mjdess...@wvhcs.org' mjdess...@wvhcs.org Message-ID: 3feff18ff4e1914a9ab7d8498591be8610cf058...@vexccr02.phsabc.ehcnet.ca Content-Type: text/plain; charset=us-ascii We use histogel a lot in our lab. It's a research lab and we use it for a few purposes - pelleting cultured cells then creating multi-culture TMAs for testing antibodies and also pelleting cells from ascites and pleural effusions. Has also been used to process really small samples that could have been lost in the processor through the cassettes. Works quite well. The researchers just put the samples in histogel and give it to me in formalin then I process it as I would regular tissue. Cuts very well too. Katy Message: 3 Date: Fri, 24 Jun 2011 12:26:46 -0400 From: Dessoye, Michael J mjdess...@wvhcs.org Subject: [Histonet] HistoGel To: histonet@lists.utsouthwestern.edu Message-ID: e2547e1cd0ee324488a2940994571efa0401f...@wvhcs-exchange.wvhcs.com Content-Type: text/plain; charset=iso-8859-1 Hello, Does anyone out there have any experience with HistoGel? It's Richard Allan/Thermo Fisher. They claim that you can embed scant tissues in the gel and then process, embed, and cut as usual. Just wondering how it works in the real world Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care System | mjdess...@wvhcs.org mailto:mjdess...@wvhcs.org | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax: 570-552-1526 -- Message: 2 Date: Fri, 24 Jun 2011 13:29:09 -0400 From: Amos Brooks amosbro...@gmail.com Subject: [Histonet] Histogel Problem To: histonet@lists.utsouthwestern.edu Message-ID: BANLkTinbTG=qcs3peuf8zfa3bz2wqrt...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Hi, I have a problem with some blocks that were prepared with Histogel. I was hoping someone else might have had a similar problem and figured it out. I took a photo of the blocks that were mads and put them in a Picassa album here: https://picasaweb.google.com/lh/photo
Re: [Histonet] Histogel Problem
Amos, I have had the same problem in the past, and posted my issues on the Histonet, however no one was able to help me out. At one point I even exchanged Histogel with a colleage severl hundred miles away, thinking that maybe I had a bad lot of Histogel. She did not have a problem with my Histogel and neither did I with her Histogel. The shrinking was arbitrary. At times all of them look great and other times more than half shrunk and looked brittle. Even RA/Thermo did not have an answer for me. I decided to do an experiment. To not bore everyone on the Histonet and explain all of my experimental steps, what it boiled down to is that you need a long processing program on the processor. We use a VIP processor as well, and the processing program is at least 12 hours long. NO MORE PROBLEMS. Since I started using this progrem, every single Histogel block has been perfect. Let me know if you need any further info or explanation. Dusko Trajkovic HT ASCP Pfizer Inc. La Jolla From: Amos Brooks amosbro...@gmail.com To: histonet@lists.utsouthwestern.edu Sent: Fri, June 24, 2011 10:29:09 AM Subject: [Histonet] Histogel Problem Hi, I have a problem with some blocks that were prepared with Histogel. I was hoping someone else might have had a similar problem and figured it out. I took a photo of the blocks that were mads and put them in a Picassa album here: https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink The long short of it is that the blocs were prepared by the researcher for me to process. They are mouse kidneys. Now it is entirely possible for him to have goofed something up in preparing the Histogel blocks. I wasn't there when he did it, but when I looked at them before processing, they all looked fine. (Like the adjacent good one in the photo). When they were processed they were placed in the VIP right next to each other. When I went to embed them this morning all but one of the four looked fine. The one that didn't come out well looked like the Histogel had shrunk up and shriveled around the kidneys. I am sure this will be aweful to cut, and the researcher is going to have a bird over it since this happened with another project previously. I would like to have a decent explanation for him, so if anyone knows what might have happened and has suggestions I would love to hear it (yes vendors too are welcome to answer this of course!). By the way, this was processed on a rather short cycle of 15-20 min per station of graded ETOH from 70% to 100% with 3 xylene stations and 4 paraffin stations (45 min for these). It seems fine for everything else that was on the processor. Just that one Histogel block was the issue. Thank you, Amos ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HistoGel
Hello, Does anyone out there have any experience with HistoGel? It's Richard Allan/Thermo Fisher. They claim that you can embed scant tissues in the gel and then process, embed, and cut as usual. Just wondering how it works in the real world Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health Care System | mjdess...@wvhcs.org mailto:mjdess...@wvhcs.org | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax: 570-552-1526 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Wyoming Valley Health Care System. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Histogel Problem
Hi, I've encountered this problem before in my previous lab. To reduce the Histogel from shrinking that badly, avoid putting the tissue at the very edge of the Histogel, space them out nicely in the middle, providing sufficient amount of extra Histogel between each tissue and surrounding them. A little bit of shrinking usually do not interfere with cutting as long as the whole thing is placed flat on the final paraffin block, the rest will stretch out on the water bath. I find sometimes the gel does not behave well in the water but as long as it is not on top of your tissue when placed on your slide, it should interfere much with staining. Rose -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Friday, June 24, 2011 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histogel Problem Hi, I have a problem with some blocks that were prepared with Histogel. I was hoping someone else might have had a similar problem and figured it out. I took a photo of the blocks that were mads and put them in a Picassa album here: https://picasaweb.google.com/lh/photo/APO3HsIMa2_jPOEs3QRGUjhz3qi22FNPb2i5JJnBCAk?feat=directlink The long short of it is that the blocs were prepared by the researcher for me to process. They are mouse kidneys. Now it is entirely possible for him to have goofed something up in preparing the Histogel blocks. I wasn't there when he did it, but when I looked at them before processing, they all looked fine. (Like the adjacent good one in the photo). When they were processed they were placed in the VIP right next to each other. When I went to embed them this morning all but one of the four looked fine. The one that didn't come out well looked like the Histogel had shrunk up and shriveled around the kidneys. I am sure this will be aweful to cut, and the researcher is going to have a bird over it since this happened with another project previously. I would like to have a decent explanation for him, so if anyone knows what might have happened and has suggestions I would love to hear it (yes vendors too are welcome to answer this of course!). By the way, this was processed on a rather short cycle of 15-20 min per station of graded ETOH from 70% to 100% with 3 xylene stations and 4 paraffin stations (45 min for these). It seems fine for everything else that was on the processor. Just that one Histogel block was the issue. Thank you, Amos ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet /prePFONT face=Verdana color=blue size=1SPAN style=FONT-SIZE: 8pt; COLOR: blue This e-mail, including any attachments, is meant only for the intended recipient and may be a confidential communication or a communication privileged by law. If you received this e-mail in error, any review, use, dissemination, distribution, or copying of this e-mail is strictly prohibited. Please notify the sender immediately of the error by return e-mail and please delete this message from your system. Thank you in advance for your cooperation. /SPAN/FONT/P ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] HistoGel for cell blocks
One of my pathologists is interested in the pros and cons of HistoGel for cell blocks. I have read the archives and gathered some information for him. He specifically would like to talk to someone that has been using it for some time. Along the same line, what method do you find to be the best for cell blocks? Any help would be appreciated. Thanks! Lynne Bell, HT (ASCP) Histology Team Leader Central Vermont Medical Center P. O. Box 547 Barre, VT 05641 802-371-4923 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] HistoGel for cell blocks
A little bit of plain agar, or a plain TSA slant tube (stolen from micro), melted in the microwave for 10 seconds, works just as well as histogel, imho. Jay A. Lundgren M.S., HTL (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] histogel technique
Next day, pellet cells and resuspend them in a small amount (100-200 microliters) of Histogel or Agar. When the gel cools cut the microcentrifuge tube open with a scalpel or razor blade. Now you have a nice cell pellet shaped like the bottom of your centrifuge tube, and you can treat it just like any piece of tissue. Pop it in a tissue processing cassette and then to your ethanol dehydrating steps. We use histogel routinely for clinical specimens. We have found several things that improve the process: 1)after you pellet the cells and decant, use a cotton tip swab to pick up the last little droplet of fluid adjacent to the button. It will hold together much better. 2)after you have added histogel, vortex and centrifuge to pellet. Put tube in ice bath, add a few drops of formalin, and wait a few minutes. With minimal prodding the plug will essentially pour out of the tube. No need to cut. Bill Tench Associate Dir. Laboratory Services Chief, Cytology Services Palomar Medical Center 555 E. Valley Parkway Escondido, California 92025 bill.te...@pph.org Voice: 760- 739-3037 Fax: 760-739-2604 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Thursday, May 13, 2010 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [BULK] Histonet Digest, Vol 78, Issue 17 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than Re: Contents of Histonet digest... Today's Topics: 1. RE: FFPE cell pellet prep (JR R) 2. xylene substitutes (Perry, Margaret) 3. RE: AMT Cover (Jones, Laura) 4. RE: AMT Cover (wanda.sm...@hcahealthcare.com) 5. RE: AMT Cover (Jennifer Anderson) 6. RE: RE: AMT Cover (Terri Brown) 7. RE: AMT Cover (Bell, Lynne) 8. RE: AMT Cover (Elliott, Rachel A.) 9. 2010 Louisiana Society for Histotechnology State Meeting - Reservation Extension! (Montina Van Meter) 10. Re: Xylene substitutes (V. Neubert) 11. Gastric biopsies (Cristi stephenson) 12. RE: Forwarding Request for Control Mehtod (Tony Henwood) 13. Re: Gastric biopsies (Brandi Higgins) 14. Re: Gastric biopsies (susanfpl...@aim.com) 15. How do I subscribe? (Michelle MacVeigh-Aloni) 16. job opening for cytology/ FISH and histology supervisor - Florida (dcoj...@tampabay.rr.com) 17. RE: Gastric biopsies (Gill, Caula A.) 18. RE: Are Histotechs considered exempt employees? (Heckford, Karen - SMMC-SF) 19. RE: Gastric biopsies (Cynthia Pyse) 20. Re: Gastric biopsies (Rene J Buesa) 21. RE: Gastric biopsies (Weems, Joyce) 22. RE: Gastric biopsies (Joyce Cline) 23. Tilapia Fish Eye (Marquisha Paul) -- Message: 1 Date: Wed, 12 May 2010 10:36:15 -0700 From: JR R rosenfeld...@hotmail.com Subject: RE: [Histonet] FFPE cell pellet prep To: histonet@lists.utsouthwestern.edu Message-ID: bay135-w681d7f121be89ca27acdcdb...@phx.gbl Content-Type: text/plain; charset=iso-8859-1 Here's how I do it. I hate the lens paper method. I transfer suspended cells from a T75 flask to a 50 ml Falcon tube and centrifuge at 400 X g for 5 minutes. Aspirate most of the media so that cells plus media are about 1 ml. Transfer cells plus media to a 1.7 ml microcentrifuge tube. Pellet cells at 400 x G for 5 minutes. Rinse with PBS if needed. Suspend cells in NBF and fix over night. Next day, pellet cells and resuspend them in a small amount (100-200 microliters) of Histogel or Agar. When the gel cools cut the microcentrifuge tube open with a scalpel or razor blade. Now you have a nice cell pellet shaped like the bottom of your centrifuge tube, and you can treat it just like any piece of tissue. Pop it in a tissue processing cassette and then to your ethanol dehydrating steps. Jerry Ricks Research Scientist University of Washington Department of Pathology Date: Wed, 12 May 2010 06:50:14 -0700 From: kmerriam2...@yahoo.com To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FFPE cell pellet prep Sorry - I forgot to put a subject line. Good morning, I am trying to make some nice FFPE cell pellet blocks, but I seem to lose a lot of cells along the way (especially when trying to take the cells out of the tube). We are fixing the cells in NBF, spinning them down, adding 70% and spinning down again. At that point, I am trying to scoop out the cells (with a weighing scoop) and wrap them in lens paper for processing. I am
[Histonet] Histogel
Hello everyone! If you are using Histogel for your cytology cell blocks, how do you liquify it? Do you use an incubator or microwave? How happy are the pathologists with the quality of the cell blocks? Thanks in advance for your information. Carol Bryant, CT (ASCP) Cytology/Histology Manager Pathology Services Lexington Clinic Phone (859) 258-4082 Fax (859) 258-4081 cb...@lexclin.com NOTICE OF CONFIDENTIALITY This message, including any attachments, is intended only for the sole use of the addressee and may contain confidential or privileged information that is protected by the State of Kentucky and/or Federal regulations. If you are not the intended recipient, do not read, copy, retain or disseminate this message or any attachment. If you have received this message in error, please call the sender immediately at (859)258-4000 and delete all copies of this message and any attachment. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. Neither the transmission of this message or any attachment, nor any error in transmission or misdelivery shall constitute waiver of any applicable legal privilege. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histogel
Wow, what is the histogrl backlash? Will someone please summarize the issues people are having? Thanks! Andrea -Original Message- From: Hofecker, Jennifer L jennifer.l.hofec...@vanderbilt.edu Date: Fri, 20 Mar 2009 08:23:02 To: histonet@lists.utsouthwestern.edu; Andrea Granthamalgra...@u.arizona.edu Subject: RE: [Histonet] processing v-e-r-y tiny samples Hi Andi, Happy Friday! I know there's been a Histogel backlash of late, but I would still recommend trying it. I haven't had any problems. We use it for scant neuro specimens every day. We also have used H.G. for mouse sural nerve biopsies that needed to retain orientation (very tiny). If you do use Histogel, do not put it in a histoscreen or biopsy cassette and be sure to put it on a process that's long enough. People often use the cassette/process based on the size of the tissue, not of the Histogel block. I'd ask thermo for a sample of H.G. and try it. If you need any Histogel help, let me know. I'm a huge fan! Have a great day, Jennifer Jennifer L. Hofecker HT(ASCP) Vanderbilt University Medical Center Division of Neuropathology Nashville, TN ph 615.343.0083 fax 615.343.7089 -Original Message- From: Bryan Watson [mailto:bryan.wat...@parkview.com] Sent: Thursday, March 19, 2009 3:39 PM To: histonet@lists.utsouthwestern.edu; Andrea Grantham Subject: Re: [Histonet] processing v-e-r-y tiny samples I may never complain about tiny GI or bronch biopsies ever again! Andrea Grantham algra...@u.arizona.edu 3/19/2009 11:51 AM Good Morning! In keeping with the weirdness of the projects I get in this lab today my question is about processing mosquito GI tracts. I have a processing schedule - that is not the problem. I'm wondering if anybody out in histoland has a suggestion for what kind of cassette to use. I was thinking of the histoscreen cassette because these GI tracts are so thin (I think thinner than a hair)and I don't want to wrap them or use sponges because I'm afraid that I'll loose them or crush them. Any ideas? Andi . : Andrea Grantham, HT(ASCP) Dept. of Cell Biology Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415)Tucson, AZ 85724-5044USA : : (FAX: 520-626-2097) (email: algra...@u.arizona.edu) : :...: http://www.cba.arizona.edu/histology-lab.html ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Histogel
histogrl backlash thats a whole 'nother set of issues altogether ;) happy friday! anh2...@med.cornell.edu wrote: Wow, what is the histogrl backlash? Will someone please summarize the issues people are having? Thanks! Andrea -Original Message- From: Hofecker, Jennifer L jennifer.l.hofec...@vanderbilt.edu Date: Fri, 20 Mar 2009 08:23:02 To: histonet@lists.utsouthwestern.edu; Andrea Granthamalgra...@u.arizona.edu Subject: RE: [Histonet] processing v-e-r-y tiny samples Hi Andi, Happy Friday! I know there's been a Histogel backlash of late, but I would still recommend trying it. I haven't had any problems. We use it for scant neuro specimens every day. We also have used H.G. for mouse sural nerve biopsies that needed to retain orientation (very tiny). If you do use Histogel, do not put it in a histoscreen or biopsy cassette and be sure to put it on a process that's long enough. People often use the cassette/process based on the size of the tissue, not of the Histogel block. I'd ask thermo for a sample of H.G. and try it. If you need any Histogel help, let me know. I'm a huge fan! Have a great day, Jennifer Jennifer L. Hofecker HT(ASCP) Vanderbilt University Medical Center Division of Neuropathology Nashville, TN ph 615.343.0083 fax 615.343.7089 -Original Message- From: Bryan Watson [mailto:bryan.wat...@parkview.com] Sent: Thursday, March 19, 2009 3:39 PM To: histonet@lists.utsouthwestern.edu; Andrea Grantham Subject: Re: [Histonet] processing v-e-r-y tiny samples I may never complain about tiny GI or bronch biopsies ever again! Andrea Grantham algra...@u.arizona.edu 3/19/2009 11:51 AM Good Morning! In keeping with the weirdness of the projects I get in this lab today my question is about processing mosquito GI tracts. I have a processing schedule - that is not the problem. I'm wondering if anybody out in histoland has a suggestion for what kind of cassette to use. I was thinking of the histoscreen cassette because these GI tracts are so thin (I think thinner than a hair)and I don't want to wrap them or use sponges because I'm afraid that I'll loose them or crush them. Any ideas? Andi . : Andrea Grantham, HT(ASCP) Dept. of Cell Biology Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415)Tucson, AZ 85724-5044USA : : (FAX: 520-626-2097) (email: algra...@u.arizona.edu) : :...: http://www.cba.arizona.edu/histology-lab.html ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histogel
Has anyone tried embedding a block of tissue like a mouse brain in Histogel, and then cutting frozen or on a vibratome? I am doing Golgi stain on mouse brain, and the tissue is VERY friable. Website says, No matter how small, friable, or viscous your histology or cytology specimen, HistoGel will encapsulate and retain the entire specimen during histological processing Bob Nienhuis UCLA / VA Medical Center On Tue, Jan 13, 2009 at 9:49 AM, Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote: Histogel works great and does not absorb water like plain agar does. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, January 13, 2009 12:46 PM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing 3-dimensional cell cultures into paraffin I have processed and embedded cells grown in/on agar, or post-embedded in agar or agarose, many times. It really isn't much different from processing and embedding tissue. You can trim the agar blocks to appropriate size with a scalpel (if necessary) either before or after fixation (if you trim them before fixation, do it while they are cold, so the agar is firm), and place them in a cassette just like a piece of tissue. Put them on the processor as usual, processing times the same as you would use for tissues of equivalent size, and embed as usual. One difference in sectioning however. Don't place the blocks in water after facing them off. The agar absorbs too much water and becomes soft. The blocks should be cut cold, but not moistened. Either place them on a cold plate without water, or put them in the refrigerator after facing, and take them out one at a time as you section them. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
