Re: [Histonet] Sections falling off
Thanks a lot! El 22/03/2017 a las 20:33, LitePath escribió: Hi Pablo This article just came out, I actually have not read yet, just downloaded yesterday...thought I would send to your way as it might be helpful... Salute Luis Luis Chiriboga PhD Director IHC Experimental Pathology NYULMC *From:* Pablo Sánchez Quinteiro via Histonet <histonet@lists.utsouthwestern.edu> *To:* histonet@lists.utsouthwestern.edu *Sent:* Wednesday, March 22, 2017 1:50 PM *Subject:* [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu <mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Thanks a lot Shirley, How long you wash in water the tissue before embedding? Where does the tissue come from? 70 alcohol? Pablo El 22/03/2017 a las 20:06, Shirley A. Powell escribió: I cut autopsy material for the medical examiners here and they are usually very bloody and thick sections. I face my blocks, soak them in an ice slush for 30 min to an hour or on a moist towel in the freezer overnight. I section them at 5 microns, a little thinner than yours. I use a product called Sta-On from Leica, it was a Surgipath product but Leica bought them out. I use a 10 ml to 990 ml water in my water bath. Charged slides or coated ones do not hold the bloody tissues on as well, so I use plain slides. After sectioning I stand them up for all the water to drain from between the section and the slide. Next I place on a hot plate/slide warmer at a temp just above the melting point of my paraffin to help the section relax and smooth out the micro folds that may be in the section. This takes s just a few minutes to melt the paraffin from around and in the section. I then place in the slide staining rack and place in an hot air slide dryer for at least an hour, but for me, I leave them overnight at 60 degrees. The next morning I stain my sections. Brain is a tissue that does not adhere as well as others to the slide so when staining be very gentle, agitate them in each solution but I would not agitate vigorously. The reason I asked about washing the bouin's out, is that it could change the charge on your slides and affect the stain. I was always taught to wash the tissue before beginning the dehydration in the processing so that residual picric acid would not affect my staining as well. Hope this makes sense and helps. Shirley -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 2:22 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sections falling off Thanks for your reply Shirley. After 24 h fixation in Bouin samples are passed into 70º ethanol. I have tried drying the sections at RT for short or long period (30´) and then 37º oven overnight. The tissue is mouse brain, sections thickness 8 microns. Pablo El 22/03/2017 a las 18:45, Shirley A. Powell escribió: Are you washing the bouins out of the tissue well before processing, that may be one source. Also how long are you drying your sections before staining? What type of tissue are you processing? -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Yes I do... if they do not fall off in the hydration. Pablo El 22/03/2017 a las 20:04, Ingles Claire escribió: Pablo Do you wash the slides in water before staining to remove the Bouin's (until the sections are no longer yellow)? I seem to remember that needs to be done. Claire From: Pablo Sánchez Quinteiro via Histonet [histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:29 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sections falling off Thanks for your hint Philip, They are stored at 37º overnight. I learnt the problem with the trapped water. I am very careful with it but they keep spoiling. They seem transparent. Could I try 60 minutes at 60º oven just before xylen? Pablo El 22/03/2017 a las 18:53, Manfre, Philip escribió: -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Pablo, I think your sections are not completely dry, after placing them on slides. Silanized slides can trap water between the sections and the slide, almost appearing like a bulge. You may need to use the corner of a paper towel to "poke" the paraffin section and drain off the excess water. Make sure the sections look dry before starting the staining procedure. They should be transparent (translucent), not opaque (milky white). That's my guess Phil. Philip Manfre, HT (ASCP) Associate Principal Scientist Merck Research Laboratories WP81-406 770 Sumneytown Pike West Point, PA 19486 215-652-9750 215-993-0383 (fax) philip_man...@merck.com Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (2000 Galloping Hill Road, Kenilworth, New Jersey, USA 07033), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet. --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Thanks for your hint Travis. The slides are new and good. Dako Flex Immuno Slides. Pablo El 22/03/2017 a las 18:43, Travis O'Brien escribió: Could be bad or expired Slides! What type are you using? Travis O’Brien International Medical Equipment, Inc. 170 Vallecitos De Oro San Marcos, CA 92069 800.543.8496 Phone 760.761.0859 Fax www.imebinc.com -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 10:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet . --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Thanks for your hint Philip, They are stored at 37º overnight. I learnt the problem with the trapped water. I am very careful with it but they keep spoiling. They seem transparent. Could I try 60 minutes at 60º oven just before xylen? Pablo El 22/03/2017 a las 18:53, Manfre, Philip escribió: -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Pablo, I think your sections are not completely dry, after placing them on slides. Silanized slides can trap water between the sections and the slide, almost appearing like a bulge. You may need to use the corner of a paper towel to "poke" the paraffin section and drain off the excess water. Make sure the sections look dry before starting the staining procedure. They should be transparent (translucent), not opaque (milky white). That's my guess Phil. Philip Manfre, HT (ASCP) Associate Principal Scientist Merck Research Laboratories WP81-406 770 Sumneytown Pike West Point, PA 19486 215-652-9750 215-993-0383 (fax) philip_man...@merck.com Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (2000 Galloping Hill Road, Kenilworth, New Jersey, USA 07033), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Thanks for your reply Jeanine, Sections are 8 microns thick (mouse brain) and they are stored overnight at 37ºC Pablo El 22/03/2017 a las 18:51, Sanders, Jeanine (CDC/OID/NCEZID) escribió: Hi Pablo. How thick are your sections and how long and at what temperature do you heat the slides before beginning de-paraffinizing? Jeanine H. Sanders Infectious Diseases Pathology Branch Centers for Disease Control and Prevention 1600 Clifton Rd., NE MS-G32 Atlanta, GA 30329 -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Sections falling off
Thanks for your reply Shirley. After 24 h fixation in Bouin samples are passed into 70º ethanol. I have tried drying the sections at RT for short or long period (30´) and then 37º oven overnight. The tissue is mouse brain, sections thickness 8 microns. Pablo El 22/03/2017 a las 18:45, Shirley A. Powell escribió: Are you washing the bouins out of the tissue well before processing, that may be one source. Also how long are you drying your sections before staining? What type of tissue are you processing? -Original Message- From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, March 22, 2017 1:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Sections falling off
Dear Histonetters, I am in trouble with my paraffin sections -samples fixed in Bouin's liquid. Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised. I do not think it was an adhesion problem. What do you think about? Embedding, cuttibg? Thanks a lot in advance Pablo --- El software de antivirus Avast ha analizado este correo electrónico en busca de virus. https://www.avast.com/antivirus ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] KU Size
Dear Histonetters, Searching for a product -an enzyme- its size it is not indicated in volume or weight. It is labelled as 25 KU. Does it mean that the vial contains 25.000 activity units? Thanks Pablo ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
