RE: [Histonet] Refurbished histology Leica equipment
We have the Leica XL autostainer and CV 5030 coverslipper, both refurbished units, and are very happy with them. We saved a lot of money by doing so. We purchased both from Medical Equipment Source. We also purchased a Leica RM2255 automatic microtome as well. Peggy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Damaris Beil Sent: Friday, November 12, 2010 6:54 PM To: histonet Subject: [Histonet] Refurbished histology Leica equipment Hello, I would like to know if anyone has had any experience using a refurbished Leica XL auto stainer and coverslip CV5000. Any thoughts you would not mind sharing? Thanks in advance, ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Tap Water or filtration system
We also use tap water, but have a filter unit attached. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Vanessa Avalos Sent: Monday, November 15, 2010 12:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tap Water or filtration system We are in the process of getting our new Lecia Autostainer delivered to us. The question came up if we will still need to continue using our water purification system for the stainer. Mostly the docs would like to save money but I don't want them to be unhappy with the result. I would like to hear all the pros and cons of discontinuing/continuing the use of our system and going w/ tap water from the sink. What is the PH level supposed to test at? What difference will I see in the slides? By the way, I only stain HE at the current time but there is a possibility of starting PAS or other special stains. Thank you in advance!!! V.Avalos ADS, INC Fax:602-277-2134 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Control sections for frozen sections
Hi Diana, We run one frozen section HE every morning after the frozen section staining reagents have been filtered, changed or topped up. The QC slide is checked by the technologist and the results recorded and initialled on the QC sheet. Lee Hamilton, Ontario ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] neurodegeneration stains?
Hi all, I am trying to characterize neuronal degeneration and was wondering if there are any fluorescent markers out there to stain for dead/dying neurons? I have used Silver stain and am currently trying to optimize the Fluro Jade C marker too. But I need some more and the literature seems limited on this. Thanks -- Anjum ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cardboard slide flat storage
Meghan, This is the catalog number for the slide/block storage boxes we use: 22-272-957 and we get them from Fisher Scientific. The following is the link on fisher's website: http://www.fishersci.com/wps/portal/PRODUCTDETAIL?prodcutdetail='prod'produ ctId=1589085catalogId=29104matchedCatNo=22272957pos=1catCode=RE_SCendec aSearchQuery=%23store%3DScientific%23N%3D0%23rpp%3D15fromCat=yeskeepSessio nSearchOutPut=truefromSearch=YsearchKey=272957||22highlightProductsItemsF lag=Y Hope that helps! Michelle __ Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cardboard slide flat storage Hi Meghan, We use grey colored boxes that will hold either 8 cardboard drawers of blocks or 4 cardboard drawers of slides. I am at home right now, don't have the specific ordering information, but will share it tomorrow when I get to work. I just didn't want to let the question languish w/o some sort of response! Michelle On Nov 15, 2010, at 9:30 AM, Meghan Tucker meghan.tuc...@yahoo.com wrote: Good morning, Are there any suggestions on a place to order a storage shelf for cardboard slide flats, which may have 6-8 'cubbies' that can be used to organize the flats? Thanks! Meghan meghan.tuc...@yahoo.com ___ 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 - No virus found in this message. Checked by AVG - www.avg.com Version: 10.0.1153 / Virus Database: 424/3260 - Release Date: 11/16/10 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Partially muddy slides
Hi Histonetters! We are having an issue in our lab with some inconsistent staining. It's an odd situation. One small biopsy can stain reasonably well over most of the tissue, but then one area of it is muddy. This does not affect all the slides in the same staining rack, nor does it affect all the small biopsies that were processed in the same processing cycle. Troubleshooting this has been difficult, at best. It's sort of hit or miss on when/where the muddy-ness appears. Here are my thoughts: ** I don't know if this is a processing issue, a staining issue or perhaps even a collection issue. ** If it affected all small biopsies equally, I would be tempted to question the processing/fixation. But it doesn't affect across the board. ** If all the staining was muddy, I would look to the stainer, but again, even within the same slide, we can have clear and muddy areas. ** The stainer is a Leica XL (about 3 years old), hooked up to tap water for the washes. ** We are using a regressive staining method with Fisher brand Protocol Harris hematoxylin and Protocol Eosin Y. We use acid alcohol and ammonia water for differentiating and bluing. ** The xylene we use is recycled, but verified for purity. ** The alcohols are recycled, but the last alcohol prior to the xylene/coverslipping is pure, not recycled (for just in case). Suggestions on what could be causing this issue would be greatly appreciated. Michelle ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Disposable blade holder for a Leica 1512 rotary microtome.
I hope that someone on histonet could help me. Our research lab just acquired an old Leica 1512 in good working order but lacking a blade holder. Is it possible for us to find a blade holder for this machine or is this a futile cause? Any information you all could provide would be greatly appreciated. Thanks, Bret Clough Texas AM Health Science Center Temple TX. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] neurodegeneration stains?
Anjum Those are the two that I'm familiar with. I'm not aware of any others. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ANJUM PARKAR Sent: Tuesday, November 16, 2010 8:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] neurodegeneration stains? Hi all, I am trying to characterize neuronal degeneration and was wondering if there are any fluorescent markers out there to stain for dead/dying neurons? I have used Silver stain and am currently trying to optimize the Fluro Jade C marker too. But I need some more and the literature seems limited on this. Thanks -- Anjum ___ 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] Partially muddy slides
Regarding incomplete deparaffinizing: I did give that some thought, but wouldn't that affect ALL the slides and not just hit or miss? It's primarily the small biopsies that are being affected, and I would think those would clear the paraffin much easier than the big specimens? Just as a troubleshooting step, I will increase the deparaffinzing times in the stainer. Jeff, I wondered if the collecting nurse/doctor was possibly placing the tissue in saline prior to formalin? A cautery effect... Would that show up as muddy or more burned/desiccated? Please keep the ideas coming! Michelle -Original Message- From: Victor Tobias [mailto:vic...@pathology.washington.edu] Sent: Tuesday, November 16, 2010 12:14 PM To: histot...@imagesbyhopper.com Subject: Re: [Histonet] Partially muddy slides What about incomplete paraffin removal? Without seeing the slides it is hard to tell. Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 vic...@pathology.washington.edu 206-598-2792 206-598-7659 Fax = Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. On 11/16/2010 9:09 AM, histot...@imagesbyhopper.com wrote: Hi Histonetters! We are having an issue in our lab with some inconsistent staining. It's an odd situation. One small biopsy can stain reasonably well over most of the tissue, but then one area of it is muddy. This does not affect all the slides in the same staining rack, nor does it affect all the small biopsies that were processed in the same processing cycle. Troubleshooting this has been difficult, at best. It's sort of hit or miss on when/where the muddy-ness appears. Here are my thoughts: ** I don't know if this is a processing issue, a staining issue or perhaps even a collection issue. ** If it affected all small biopsies equally, I would be tempted to question the processing/fixation. But it doesn't affect across the board. ** If all the staining was muddy, I would look to the stainer, but again, even within the same slide, we can have clear and muddy areas. ** The stainer is a Leica XL (about 3 years old), hooked up to tap water for the washes. ** We are using a regressive staining method with Fisher brand Protocol Harris hematoxylin and Protocol Eosin Y. We use acid alcohol and ammonia water for differentiating and bluing. ** The xylene we use is recycled, but verified for purity. ** The alcohols are recycled, but the last alcohol prior to the xylene/coverslipping is pure, not recycled (for just in case). Suggestions on what could be causing this issue would be greatly appreciated. Michelle ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - No virus found in this message. Checked by AVG - www.avg.com Version: 10.0.1153 / Virus Database: 424/3260 - Release Date: 11/16/10 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] connexin 43 ab
Dear Histonetters Does anybody have a preference for a connexin-43 ab? A registrar needs it for a research project. I am leaning towards the Sigma C6219 rabbit plyclonal. I would appreciate any suggestions. The material would be ffpe human heart tissue Many thanks Heather McLeod South Africas premier free email service - www.webmail.co.za Save on insurance with OUTsurance https://www.outsurance.co.za/insurance-quote/?source=webmailmailercr=thou10_468x60cid=37 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Vol 84, Issue 20 Non-Refrigerated RPMI/3-in-1 Pap Stain
For those who were in Seattle this is the info you are looking for ... The RPMI storage temp is +15 C to + 30 C and we purchase ours from Fisher (cat# 12-167F Lonza BioWhittaker RPMI-1640 500ml www.fishersci.comhttp://www.fishersci.com/ The 3-in-1 Pap is called EasyPap stain and we purchase ours from Azer scientific cat# ES7037A www.AzerSci.comhttp://www.azersci.com/ Bill Lecorchick Cytology Prep.Tech. 609-584-5128 Fax 609-584-6439 wleco...@rwjuhh.edu www.rwjhamilton.orghttp://www.rwjhamilton.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FW: What % of IHC Stains Should You Expect to Repeat?
WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _ From: Smith Wanda Sent: Tuesday, November 16, 2010 1:59 PM To: 'histonet-requ...@lists.utsouthwestern.edu' Subject: What % of IHC Stains Should You Expect to Repeat? Good Afternoon, I am having an issue with my Pathologist who thinks when the patient tissue on an IHC stain doesn't look like what he thought it should look like, he then wants to stop doing Immunos and send everything out This may happen about twice a month, but the control always stains beautifully. We recently had the stainer serviced and the drop zone was adjusted to alleviate any problems with distribution of the solutions. On a recent test slide of a new control block of colon cancer, he also didn't like that CD X2 did not stain some of the tissue components, but the rest of the tissue on the test slide stained beautifully. Please advise me on what to say to this Pathologist. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Partially muddy slides
Is it the same person cutting all of the problem biopsies? Does your lab use cryo spray? We had biopsies that had a muddy/freezer burnt appearance. We traced all of the problems to one histotech who used the cryo spray excessively. histot...@imagesbyhopper.com Sent by: histonet-boun...@lists.utsouthwestern.edu 11/16/2010 09:12 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Partially muddy slides Hi Histonetters! We are having an issue in our lab with some inconsistent staining. It's an odd situation. One small biopsy can stain reasonably well over most of the tissue, but then one area of it is muddy. This does not affect all the slides in the same staining rack, nor does it affect all the small biopsies that were processed in the same processing cycle. Troubleshooting this has been difficult, at best. It's sort of hit or miss on when/where the muddy-ness appears. Here are my thoughts: ** I don't know if this is a processing issue, a staining issue or perhaps even a collection issue. ** If it affected all small biopsies equally, I would be tempted to question the processing/fixation. But it doesn't affect across the board. ** If all the staining was muddy, I would look to the stainer, but again, even within the same slide, we can have clear and muddy areas. ** The stainer is a Leica XL (about 3 years old), hooked up to tap water for the washes. ** We are using a regressive staining method with Fisher brand Protocol Harris hematoxylin and Protocol Eosin Y. We use acid alcohol and ammonia water for differentiating and bluing. ** The xylene we use is recycled, but verified for purity. ** The alcohols are recycled, but the last alcohol prior to the xylene/coverslipping is pure, not recycled (for just in case). Suggestions on what could be causing this issue would be greatly appreciated. Michelle ___ 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] Biocare¹s Rodent Decloaking Solution
Hi everyone, For anyone that uses Biocare's Rodent Decloaking Solution, once the solution has been made up, can it be reused? And if so in your experience how many times? Many thanks in advance ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] FW: What % of IHC Stains Should You Expect to Repeat?
Send the colon cancer control out and compare your results to the send out. If they are the same then you have an argument. From: wanda.sm...@hcahealthcare.com To: histonet@lists.utsouthwestern.edu Date: Tue, 16 Nov 2010 13:07:28 -0600 Subject: [Histonet] FW: What % of IHC Stains Should You Expect to Repeat? WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _ From: Smith Wanda Sent: Tuesday, November 16, 2010 1:59 PM To: 'histonet-requ...@lists.utsouthwestern.edu' Subject: What % of IHC Stains Should You Expect to Repeat? Good Afternoon, I am having an issue with my Pathologist who thinks when the patient tissue on an IHC stain doesn't look like what he thought it should look like, he then wants to stop doing Immunos and send everything out This may happen about twice a month, but the control always stains beautifully. We recently had the stainer serviced and the drop zone was adjusted to alleviate any problems with distribution of the solutions. On a recent test slide of a new control block of colon cancer, he also didn't like that CD X2 did not stain some of the tissue components, but the rest of the tissue on the test slide stained beautifully. Please advise me on what to say to this Pathologist. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. ___ 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] Disposable blade holder for a Leica 1512 rotary microtome.
Klaus Dern at 706-635-8840 will make you one that works like a regular knife holding a disposable blade. I copied everyone in case someone else needs the info. J Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Clough, Bret Sent: Tuesday, November 16, 2010 12:10 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Disposable blade holder for a Leica 1512 rotary microtome. I hope that someone on histonet could help me. Our research lab just acquired an old Leica 1512 in good working order but lacking a blade holder. Is it possible for us to find a blade holder for this machine or is this a futile cause? Any information you all could provide would be greatly appreciated. Thanks, Bret Clough Texas AM Health Science Center Temple TX. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] FW: What % of IHC Stains Should You Expect to Repeat?
That type of pathologist deserves an answer that could cost you your job, so it is not advisable to do that. That type of pathologist is essentially ignorant of what we do and do not reason. If he wants to send everything out, let him. When charges begin to hit your administration, perhaps he will be more open to reasoning. René J. --- On Tue, 11/16/10, wanda.sm...@hcahealthcare.com wanda.sm...@hcahealthcare.com wrote: From: wanda.sm...@hcahealthcare.com wanda.sm...@hcahealthcare.com Subject: [Histonet] FW: What % of IHC Stains Should You Expect to Repeat? To: histonet@lists.utsouthwestern.edu Date: Tuesday, November 16, 2010, 2:07 PM WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _ From: Smith Wanda Sent: Tuesday, November 16, 2010 1:59 PM To: 'histonet-requ...@lists.utsouthwestern.edu' Subject: What % of IHC Stains Should You Expect to Repeat? Good Afternoon, I am having an issue with my Pathologist who thinks when the patient tissue on an IHC stain doesn't look like what he thought it should look like, he then wants to stop doing Immunos and send everything out This may happen about twice a month, but the control always stains beautifully. We recently had the stainer serviced and the drop zone was adjusted to alleviate any problems with distribution of the solutions. On a recent test slide of a new control block of colon cancer, he also didn't like that CD X2 did not stain some of the tissue components, but the rest of the tissue on the test slide stained beautifully. Please advise me on what to say to this Pathologist. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. ___ 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] Partially muddy slides
Michelle, It might possible be water contamination in the xylene. Try increasing alcohol dehydration times. Also re-cyled alcohol is probably around 96% pure. It seems to be not possible to obtain absolute ethanol by re-cycling, or so I am lead to believe. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Wednesday, 17 November 2010 4:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Partially muddy slides Hi Histonetters! We are having an issue in our lab with some inconsistent staining. It's an odd situation. One small biopsy can stain reasonably well over most of the tissue, but then one area of it is muddy. This does not affect all the slides in the same staining rack, nor does it affect all the small biopsies that were processed in the same processing cycle. Troubleshooting this has been difficult, at best. It's sort of hit or miss on when/where the muddy-ness appears. Here are my thoughts: ** I don't know if this is a processing issue, a staining issue or perhaps even a collection issue. ** If it affected all small biopsies equally, I would be tempted to question the processing/fixation. But it doesn't affect across the board. ** If all the staining was muddy, I would look to the stainer, but again, even within the same slide, we can have clear and muddy areas. ** The stainer is a Leica XL (about 3 years old), hooked up to tap water for the washes. ** We are using a regressive staining method with Fisher brand Protocol Harris hematoxylin and Protocol Eosin Y. We use acid alcohol and ammonia water for differentiating and bluing. ** The xylene we use is recycled, but verified for purity. ** The alcohols are recycled, but the last alcohol prior to the xylene/coverslipping is pure, not recycled (for just in case). Suggestions on what could be causing this issue would be greatly appreciated. Michelle ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * 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 are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Frozen brain slices
Thanks Tina. I will try that for the already frozen brains. Also I did some slicing today with some of my frozen brains and kept the block in the cryostat till it came to temperature and as compared to before I think I can say that the slices came out pretty well. I guess the temperature is the trick here. Well I am going to run some stains on them and see how they turn out and then I can say for sure that I have a good working protocol hopefully. These slices are for slide mounting not free floating. I do use the anti-roll plate and I did realize that it was a little off in its angle with respect to the knife and hence one another issue that needed fixing. Will let you on my progress. Thanks Liz for letting me know. Have you tried the Fluro Jade? The original, B or C? On Tue, Nov 16, 2010 05:33 PM, Montina Van Meter montina.vanme...@pbrc.edu wrote: Anjum, The post-fix and sucrose are for the new harvests. Do you free-float the sections or mount them on slides? Make sure you allow the -80C block of tissue to sit in the cryostat for about 20 min. to allow the block to come up to the -20C temperature of the cryostat chamber. There is no such thing as a too hard block. Allowing it to become the same temperature as the inside of the cryostat will solve that problem. Warming up the block after freezing, than freezing it down again, is an absolute no-no. This will also cause freeze/thaw artifact. Are you using an anti-roll plate when sectioning on the cryostat? That might help flatten out the sections as they come off the knife edge. If you haven't already frozen the other brains I would place them in 20% sucrose/PBS overnight. The next morning rinse off the sucrose with PBS for about 30 min. Freeze the brain on dry ice in the OCT. If you have already frozen all of the brains, you might take one of them and thaw it out. Place it in the sucrose overnight and see if that helps with the cutting. It can't hurt, since you are already having trouble getting sections. Good luck, Tina Montina J. Van Meter Lab Manager Autonomic Neuroscience 225-763-2564 vanme...@pbrc.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of ANJUM PARKAR Sent: Monday, November 15, 2010 9:24 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Frozen brain slices Thanks Tina. I will keep in mind the thickness for staining purposes that is once the slicing protocol gets optimized. So we use 200-300 ml of both PBS and PFA for our perfusion. So the post fix and sucrose treatment you are suggesting, are you suggesting for the new harvests alone which I will be doing or the existing frozen perfused ones? Anjum On Sun, Nov 14, 2010 10:35 PM, Montina Van Meter montina.vanme...@pbrc.edu wrote: Anhum, How much PBS and Paraformaldehyde do you put through the (rat or mouse). I flush a perfuse our rats with 150-200ml. of PBS and 500ml. of Para. Postfix for 2 hours and place in 20% sucrose overnight until the tisue sinks to the bottom of the vial. The morphology of your tissue is going to be compromised due to freeze artifact (lack of cryoprotectant) and there will be holes in your sections. Actually, 30um thick sections are considered quite thick and are typically used for free floating techniques. Sections that are mounted on slides before IHC staining are much thinner (3-10um). I usually cut my tissue between 30-40um and manually free float the sections for IHC or IF. Tina Sent from my iPhone On Nov 14, 2010, at 9:05 PM, ANJUM PARKAR axp...@psu.edu wrote: Thanks Tina. I will keep the sucrose suggestion in mind for future harvests. For now I need to figure out how to slice the already fixed frozen tissue. Will try to equilibrate in the cryostat itself as against room temperature and see how that goes. The knife angle is 13 degrees and recently sharpened and thickness of slices 30 microns which I believe is standard for sectioning. I have sectioned previously a lot so doing hands on is something I am familiar with. But I did realize every instrument is different and while doing a procedure it always needs to be optimized until it can become routine. Will let you know how things go. Anjum On Sun, Nov 14, 2010 02:21 PM, Montina Van Meter montina.vanme...@pbrc.edu wrote: Anhum, 1. You MUST cryoprotect the fixed tissue in 15-30% sucrose (until it sinks) prior to cutting frozen sections. 2. Allow the -80C embedded tissue block to equilabrate to -20C in the cryostat for 15-20min. 3. Check the knife angle. Thickness? 4. Change knife or sharpen and make sure all screws are tight. 5. Freez/thawing of block is not a good idea (especially since it's not cryoprotected). You are introducing ice crystal artifact. 6. Check around your department (or Histology Core) to see if someone can instruct you on using the cryostat. It's always better to
Re: [Histonet] Disposable blade holder for a Leica 1512 rotary microtome.
Bert (Embedded image moved to file: pic30695.jpg) 14036833012 : High profile blade rail (twin) - set 14036833013 : Low profile blade rail (twin) - set Hope this helps Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist/Trainer Leica Microsystems Biosystems Division Technical Assistance Center 800 248 0123 x7841 847 236 3063 fax mari.ann.mailh...@leica-microsystems.com www.leica-microsystems.com Clough, Bret clo...@medicine. tamhsc.eduTo Sent by: histonet@lists.utsouthwestern.edu histonet-bounces@ histonet@lists.utsouthwestern.edu lists.utsouthwest cc ern.edu Subject [Histonet] Disposable blade holder 11/16/2010 11:09 for a Leica 1512 rotary microtome. AM I hope that someone on histonet could help me. Our research lab just acquired an old Leica 1512 in good working order but lacking a blade holder. Is it possible for us to find a blade holder for this machine or is this a futile cause? Any information you all could provide would be greatly appreciated. Thanks, Bret Clough Texas AM Health Science Center Temple TX. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email _ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Current issues with Printmates and Checkmates
Hello? Is anyone having issues with the Checkmate and/or Printmates? What is the opinion out there about using the hot foil tape methodology? Is the tape chemically resistant? Histopathy ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
