[Histonet] Affordable microtome knife reconditioning in the EC?
Hi all, I have some large (25 - 30cm) type B, C and D, both steel and tungsten carbide, microtome knives that need sharpening and/or reconditioning. Does anyone knows a company in Europe (preferably within the EC) that does that kind of work at an affordable price? By the way: what is a *normal* price for those things? Impossible to answer question, I suppose... Thanks in advance! Yvan. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Replacement glass plates for Reichert Microtome Knife Sharpener 903?
Hi all, It struck me that all those microtome knife sharpeners (AO 935, Reichert 903, AO 903...) all look very much the same. Are the glass plates interchangeable? Would the glass plates for the current Leica sharpener SP9000 (part.no. 14041819698, dimensions 292mm x 127mm x 6 mm) fit the Reichert Microtome Knife Sharpener 903? Thanks in advance for your answers! Yvan. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Affordable microtome knife reconditioning in the EC?
I don't know about Europe, but here in the US there are a few vendors that provide these services (Dorn and Hart Microedge, Sturkey, and Delaware Diamond Knives). I am not sure of the pricing for all three of these vendors, but for the past 11 years I have sent my D-profile rotary wedge and sledge/polycut tungsten-carbide knives for re- sharpening services to Dorn and Hart Microedge. They are located near Chicago, Illinois and their prices are very reasonable. Maybe you can google all three vendors for information if you get in a bind, but I am pretty sure they provide services outside the US. Jack On Apr 21, 2009, at 5:14 AM, yvan lindekens yvan_lindek...@yahoo.com wrote: Hi all, I have some large (25 - 30cm) type B, C and D, both steel and tungsten carbide, microtome knives that need sharpening and/or reconditioning. Does anyone knows a company in Europe (preferably within the EC) that does that kind of work at an affordable price? By the way: what is a *normal* price for those things? Impossible to answer question, I suppose... Thanks in advance! Yvan. ___ 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] melanoma markers on mouse
Hi All, I am trying to perform IHC for melanoma markers on mouse tissue. In particular, does anyone have hands-on experience with antibodies against Pmel17/Silv/gp100 (recognized by HMB-45 antibody in humans), Melan-A/MART-1, or MITF on mouse? Thanks Sam Perry Research Technician DFCI, Boston MA 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] Cloudy MMA embedded block
Ooi, Somehow you are getting moisture/water mixed in with the MMA solution(s). I only remember visiting the IMCB facility, so if this is not your facility then I am not sure of your laboratory environment (air temperature/humidity). The best thing I can tell you at this point is to try and find ways to reduce the chance for moisture accumulation. I am pretty sure that at a minimum you are storing your catalyst and monomer (MMA) at 4C and your softner (DBP) at room temp. Maybe you should take extra steps to ensure your chemicals are at room temp before any use. Lastly, your polymerization is taking too long for your small 20mL vials. My specimens of similar size require only 3-5 days to completely polymerize. Now, I dont remember your concentration of catalyst, but if it is around 2.5g/1L embedding solution, try skipping the 4C step and use room temp (23C) until the specimens are polymerized. Then finish them off in the oven when they are tacky or mostly hard on top. The 4C step is mostly to slow the polymerization for larger specimens requiring a larger volume of solution and/or when the lab temp cannot be controlled routinely at or below 23C. The water bath then also helps to regulate the temperature of the polymerization in the case where the lab environment is too cool (20C) and retarding the rate of polymerzation. The main thing to keep in mind is that the lab environment pretty much controls the process and is probably the main variable that will contrast labs that perform the exact same method. Jack On Apr 21, 2009, at 12:33 AM, ooi.ting.h...@nhc.com.sg wrote: Hi, I am trying very hard to get a clear MMA embedded block. I am appreciate if there is any advice on doing it. I am using 100% MMA (liquid) and 0.5% Perkadox 16 (powder) to do the embedding. I have tried to embed the sample in different conditions. However, it seems to be extremely hard to get a clear embedded block. Below are the methods that I have tried. 1. Before embedding, bring the MMA and perkadox 16 to room temperature. After a few mix it by invertion, I aliquot the solution to the glass vials that contain samples or simply glass vials that without sample. I put the glass vials (both with and without sample) in 4 degree, room temperature and even 37degree oven. 2. Mix the MMA and perkadox when it is cold, invert it a few times. Aliquot it to glass vials that contain sample or without sample. Put in 4degree, room temperature and 37degree oven. I have also tried to speed up the polymerization process by using vacumm for a few samples... Throughout all the sample that I have, I only manage to get a clear empty glass vial which I put in the 4degree for one month and a clear embedded sample which I put in 4 degree overnight and bring it out to room temperature. The rest of the glass vials (with or without sample) are cloudy. I have repeated the methods that I used to get a clear embedded block, I did not get any clear block (for both with and without sample). I am glad if there is any expert advice or guidance. We need a clear embedded sample. Thank you very much! Regards, Ooi _ Confidential information. Unauthorized use or disclosure prohibited. Refer http://www.singhealth.com.sg/ContactUs/#Disclaimer ___ 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] Cloudy MMA embedded block
Hi, We don't store our MMA or DBP in the refirgerator as the movement between 4C and room temperature can cause condensation in the bottles. This just adds water to the material and as it is cooled and re-warmed you are adding more each time and in both directions. When it comes to room temperature and then is replaced in the refrigerator it again will develop condensation. Over time the MMA will have enough water in it to become discolored. I am surprised you are not seeing the discoloration in the MMA liquid as it poured as a warning not to use it. We have fair control of temperature in the laboratory and are generally between 68F to 75F with the average as around 70F. We have never had a problem with the MMA polymerizing or developing a water content. It is stored in a dark cabinet that is in an area I would say is a little cooler than the room. We generally don't polymerize in the cold at 4C however, you timing seems long for the process. It is best not to disturb the vials too much once you are in the embedding phase. Movement or inversion of the vials can cause a problem as the MMA will polymerize from the bottom up and remixing it can slow the process. Also make sure you tops are tight at this step. Pam Marcum UPENN Vet Sch New Botlon Center - Original Message - From: Jack Ratliff ratliffj...@hotmail.com To: Ooi Ting Huay ooi.ting.h...@nhc.com.sg, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Tuesday, April 21, 2009 9:24:29 AM GMT -05:00 US/Canada Eastern Subject: Re: [Histonet] Cloudy MMA embedded block Ooi, Somehow you are getting moisture/water mixed in with the MMA solution(s). I only remember visiting the IMCB facility, so if this is not your facility then I am not sure of your laboratory environment (air temperature/humidity). The best thing I can tell you at this point is to try and find ways to reduce the chance for moisture accumulation. I am pretty sure that at a minimum you are storing your catalyst and monomer (MMA) at 4C and your softner (DBP) at room temp. Maybe you should take extra steps to ensure your chemicals are at room temp before any use. Lastly, your polymerization is taking too long for your small 20mL vials. My specimens of similar size require only 3-5 days to completely polymerize. Now, I dont remember your concentration of catalyst, but if it is around 2.5g/1L embedding solution, try skipping the 4C step and use room temp (23C) until the specimens are polymerized. Then finish them off in the oven when they are tacky or mostly hard on top. The 4C step is mostly to slow the polymerization for larger specimens requiring a larger volume of solution and/or when the lab temp cannot be controlled routinely at or below 23C. The water bath then also helps to regulate the temperature of the polymerization in the case where the lab environment is too cool (20C) and retarding the rate of polymerzation. The main thing to keep in mind is that the lab environment pretty much controls the process and is probably the main variable that will contrast labs that perform the exact same method. Jack On Apr 21, 2009, at 12:33 AM, ooi.ting.h...@nhc.com.sg wrote: Hi, I am trying very hard to get a clear MMA embedded block. I am appreciate if there is any advice on doing it. I am using 100% MMA (liquid) and 0.5% Perkadox 16 (powder) to do the embedding. I have tried to embed the sample in different conditions. However, it seems to be extremely hard to get a clear embedded block. Below are the methods that I have tried. 1. Before embedding, bring the MMA and perkadox 16 to room temperature. After a few mix it by invertion, I aliquot the solution to the glass vials that contain samples or simply glass vials that without sample. I put the glass vials (both with and without sample) in 4 degree, room temperature and even 37degree oven. 2. Mix the MMA and perkadox when it is cold, invert it a few times. Aliquot it to glass vials that contain sample or without sample. Put in 4degree, room temperature and 37degree oven. I have also tried to speed up the polymerization process by using vacumm for a few samples... Throughout all the sample that I have, I only manage to get a clear empty glass vial which I put in the 4degree for one month and a clear embedded sample which I put in 4 degree overnight and bring it out to room temperature. The rest of the glass vials (with or without sample) are cloudy. I have repeated the methods that I used to get a clear embedded block, I did not get any clear block (for both with and without sample). I am glad if there is any expert advice or guidance. We need a clear embedded sample. Thank you very much! Regards, Ooi
[Histonet] rulers
Thanks to everyone that has given me feedback on the prostate situation. I am testing a shorter run today and will hope that solves the issue. I was wondering if anyone (vendors included) knew of anywhere I could get some of the cheap plastic rulers (the kind they give away usually) that are flexible. I use them to measure my skin lesions and I had a few that seem to have walked away. I am using my scalpel handle currently to measure (in cm) but would really love some of the plastic ones. Thanks and have a great day. Jodi Jodi Putnam (HT,ASCP) Graves Gilbert Clinic Pathology Department 201 Park Street Bowling Green, KY 42102 (270) 393-2728 (voicemail) (270) 393-2795 Fax : (270) 393-2736 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 Graves-Gilbert Clinic. 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
[Histonet] Vectashield mounting media problem?
Hello everyone, Our lab has been using 'Vectashield mounting media w/ DAPI' for many years and had no problem of fading. Recently, two bottles with different lot numbers in a row gone bad in around four months (the signal disappears in front of your eyes, tested on a couple of different microscopes). 1) Has anyone experienced same problem? Vector lab only says that this is a very stable product and should not be happening when stored at 4C. 2) Can anyone recommend a good aqueous mounting media with DAPI for immunofluorescence? Thank you in advance, Regards, Suh ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Vectashield mounting media problem?
ProLong Gold w/ DAPI (Invitrogen/Molecular Probes) This is a permanent mounting media - works great! Montina J. Van Meter, HT (ASCP) Lab Manager Autonomic Neuroscience Pennington Biomedical Research Center 6400 Perkins Rd. Baton Rouge, LA 70791 225-763-2564 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Suhyoung Jeong Sent: Tuesday, April 21, 2009 9:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Vectashield mounting media problem? Hello everyone, Our lab has been using 'Vectashield mounting media w/ DAPI' for many years and had no problem of fading. Recently, two bottles with different lot numbers in a row gone bad in around four months (the signal disappears in front of your eyes, tested on a couple of different microscopes). 1) Has anyone experienced same problem? Vector lab only says that this is a very stable product and should not be happening when stored at 4C. 2) Can anyone recommend a good aqueous mounting media with DAPI for immunofluorescence? Thank you in advance, Regards, Suh ___ 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] Vectashield mounting media problem?
We had the EXACT same problem about a year ago! We don't believe we had storage issues, either. We've been using SlowFade Gold (Molecular Probes, Invitrogen) for about a year now and it's been a great aqueous mounting medium. We add our own DAPI (2ul of a 1mg/ml DAPI solution per 500ul SlowFade) and it works fine. Merced --On Tuesday, April 21, 2009 10:51 AM -0400 Suhyoung Jeong suhyoung.je...@gmail.com wrote: Hello everyone, Our lab has been using 'Vectashield mounting media w/ DAPI' for many years and had no problem of fading. Recently, two bottles with different lot numbers in a row gone bad in around four months (the signal disappears in front of your eyes, tested on a couple of different microscopes). 1) Has anyone experienced same problem? Vector lab only says that this is a very stable product and should not be happening when stored at 4C. 2) Can anyone recommend a good aqueous mounting media with DAPI for immunofluorescence? Thank you in advance, Regards, Suh ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 Biomedical Research Building School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Reminder - California Society for Histotechnology Annual Symposium
Just a reminder, registration deadline for the upcoming California Society for Histotechnology Annual Symposium/Convention is May 1st. This is a 3.5 day education forum on May 14-17 and will be held at the Westin San Francisco Airport. For further information, please see our website at: www.californiahistology.org or contact: Shirley Chu at 408-747-3765 or by email at: shirley@moldev.com. Cut-off date for hotel registration is April 22nd (online hotel registration: http://www.starwoodmeeting.com/StarGroupsWeb/res?id=0806209635key=F3921 ) Shirley Chu Application Scientist, Arcturus LCM Products Molecular Devices (now a part of MDS Analytical Technologies) 408-747-3765 | www.moleculardevices.com This e-mail and any files transmitted with it may contain privileged and/or confidential information and may be read or used only by the intended recipient. If you are not the intended recipient of the e-mail or any of its attachments, please be advised that you have received this e-mail in error and any use, dissemination, distribution, forwarding, printing or copying of this e-mail or any attached files is strictly prohibited. If you have received this e-mail in error, please immediately purge it and all attachments and notify the sender by reply e-mail or contact the sender at the number listed. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Peloris Tissue Processor
You cannot use recycled Formula 83 in the Peloris. The threshold is 88%. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of thisis...@aol.com Sent: Monday, April 20, 2009 10:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Peloris Tissue Processor Does anyone use recycled and/or fresh Formula 83 as a clearant on their Peloris Tissue Processor?? If so, can you tell me what final threshold percentage you use. Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * CONFIDENTIALITY NOTICE * This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Reusing specimen containers
Hi Histoland! I'm just wondering what folks out there are doing about specimen containers. Is anyone cleaning them out and reusing them? Is there any CAP or Joint Commission regulations that say that all specimen containers can only be used once or can you reuse them as long as they have been thoroughly cleaned? Thanks for you all your input, Kelly _ Windows Live™ SkyDrive™: Get 25 GB of free online storage. http://windowslive.com/online/skydrive?ocid=TXT_TAGLM_WL_skydrive_042009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Peloris Tissue Processor
Hello Joyce, Thank you for addressing this. You are correct, it is not recommended to use recycled clearing reagents on the Peloris; recycled alcohol is the only recycled reagent acceptable for use on Peloris. Kindest regards, James Chalmers, MLT (ASCP), MT (HHS) Applications Specialist/Trainer Leica Microsystems - Biosystems Division 2345 Waukegan Road Bannockburn, IL 60015 Phone: (847)405-5431 Fax: (847)405-7945 Joyce Cline jcl...@wchsys.or g To Sent by: Histonet histonet-bounces@ histonet@lists.utsouthwestern.edu lists.utsouthwest cc ern.edu Subject RE: [Histonet] Peloris Tissue 04/21/2009 11:32 Processor AM You cannot use recycled Formula 83 in the Peloris. The threshold is 88%. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of thisis...@aol.com Sent: Monday, April 20, 2009 10:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Peloris Tissue Processor Does anyone use recycled and/or fresh Formula 83 as a clearant on their Peloris Tissue Processor?? If so, can you tell me what final threshold percentage you use. Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * CONFIDENTIALITY NOTICE * This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. ___ 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] BRDU, Tetracycline, calcein in bone
I would recommend fixing the bones in 70% ethanol, embedding in methylmethacrylate undecalcified and cutting 8 - 10 micron thick sections for visualization of tetracycline and/or calcein labels. Decalcification will remove fluorescent labels bound to the mineral in bone as mineral is removed by decal procedures. You may need a larger microtome (Polycut) to cut these bones depending on the size. The fluorescent labels in combination with a nice von Kossa stain would certainly show formation/ mineralization of bone in this area which you can quantify using an Osteomeasure or other system, however, you may want to do a few in paraffin for optimal BRDU label preservation. I would recommend a slow embedding method for these bones - you can check our article in the JOH, June 2004, Vol. 27(2) pp119-130 which describes our slow embedding method for rat bones which can be adapted for rabbit bones. You can contact me for suggestions and with questions if you would like to. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Beta F1 antibody
Martha, I think you want the TCR-BF1 antibody. It is a T cell recptor for Beta F1. We get it from Pierce (Endogen), cat# TCR1151. We run it on the Bondmax and it works well. Jo Mauger Martha Ward mw...@wfubmc.edu 04/21/09 12:52 PM I have had a request for this antibody and have had some difficulty locating a vendor for it. We will be using the Leica Bond Max and if anyone has any tips, vendor, etc. I would appreciate some advice. Thanks in advance for your help. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center Winston-Salem, NC ___ 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] Cloudy MMA embedded block
In our lab we do store our MMA solutions in a flammable storage refrigerator per EHS requirements; however we put Molecular Sieves in all solutions to keep moisture from contaminating them. We rarely have any problems with cloudy blocks. We order them from VWR, catalog # JT2708-1, 500 g, ~$40 each; manufacturer is J.T. Baker. They last a very long time and are useful for any open non-aqueous solution. Just add a few (6-12) to each container. Hope this helps! Lori -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: Tuesday, April 21, 2009 6:45 AM To: Jack Ratliff Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cloudy MMA embedded block Hi, We don't store our MMA or DBP in the refirgerator as the movement between 4C and room temperature can cause condensation in the bottles. This just adds water to the material and as it is cooled and re-warmed you are adding more each time and in both directions. When it comes to room temperature and then is replaced in the refrigerator it again will develop condensation. Over time the MMA will have enough water in it to become discolored. I am surprised you are not seeing the discoloration in the MMA liquid as it poured as a warning not to use it. We have fair control of temperature in the laboratory and are generally between 68F to 75F with the average as around 70F. We have never had a problem with the MMA polymerizing or developing a water content. It is stored in a dark cabinet that is in an area I would say is a little cooler than the room. We generally don't polymerize in the cold at 4C however, you timing seems long for the process. It is best not to disturb the vials too much once you are in the embedding phase. Movement or inversion of the vials can cause a problem as the MMA will polymerize from the bottom up and remixing it can slow the process. Also make sure you tops are tight at this step. Pam Marcum UPENN Vet Sch New Botlon Center - Original Message - From: Jack Ratliff ratliffj...@hotmail.com To: Ooi Ting Huay ooi.ting.h...@nhc.com.sg, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Tuesday, April 21, 2009 9:24:29 AM GMT -05:00 US/Canada Eastern Subject: Re: [Histonet] Cloudy MMA embedded block Ooi, Somehow you are getting moisture/water mixed in with the MMA solution(s). I only remember visiting the IMCB facility, so if this is not your facility then I am not sure of your laboratory environment (air temperature/humidity). The best thing I can tell you at this point is to try and find ways to reduce the chance for moisture accumulation. I am pretty sure that at a minimum you are storing your catalyst and monomer (MMA) at 4C and your softner (DBP) at room temp. Maybe you should take extra steps to ensure your chemicals are at room temp before any use. Lastly, your polymerization is taking too long for your small 20mL vials. My specimens of similar size require only 3-5 days to completely polymerize. Now, I dont remember your concentration of catalyst, but if it is around 2.5g/1L embedding solution, try skipping the 4C step and use room temp (23C) until the specimens are polymerized. Then finish them off in the oven when they are tacky or mostly hard on top. The 4C step is mostly to slow the polymerization for larger specimens requiring a larger volume of solution and/or when the lab temp cannot be controlled routinely at or below 23C. The water bath then also helps to regulate the temperature of the polymerization in the case where the lab environment is too cool (20C) and retarding the rate of polymerzation. The main thing to keep in mind is that the lab environment pretty much controls the process and is probably the main variable that will contrast labs that perform the exact same method. Jack On Apr 21, 2009, at 12:33 AM, ooi.ting.h...@nhc.com.sg wrote: Hi, I am trying very hard to get a clear MMA embedded block. I am appreciate if there is any advice on doing it. I am using 100% MMA (liquid) and 0.5% Perkadox 16 (powder) to do the embedding. I have tried to embed the sample in different conditions. However, it seems to be extremely hard to get a clear embedded block. Below are the methods that I have tried. 1. Before embedding, bring the MMA and perkadox 16 to room temperature. After a few mix it by invertion, I aliquot the solution to the glass vials that contain samples or simply glass vials that without sample. I put the glass vials (both with and without sample) in 4 degree, room temperature and even 37degree oven. 2. Mix the MMA and perkadox when it is cold, invert it a few times. Aliquot it to glass vials that contain sample or without sample. Put in 4degree, room temperature and 37degree oven. I have also tried to speed up the
Re: [Histonet] BRDU, Tetracycline, calcein in bone
You can also perform BrdU on the MMA sections the BrdU label is quite hardy. Robert Schoonhoven From: Nancy W. Troiano nancy.troi...@yale.edu To: histonet@lists.utsouthwestern.edu Sent: Tuesday, April 21, 2009 1:37:01 PM Subject: [Histonet] BRDU, Tetracycline, calcein in bone I would recommend fixing the bones in 70% ethanol, embedding in methylmethacrylate undecalcified and cutting 8 - 10 micron thick sections for visualization of tetracycline and/or calcein labels. Decalcification will remove fluorescent labels bound to the mineral in bone as mineral is removed by decal procedures. You may need a larger microtome (Polycut) to cut these bones depending on the size. The fluorescent labels in combination with a nice von Kossa stain would certainly show formation/ mineralization of bone in this area which you can quantify using an Osteomeasure or other system, however, you may want to do a few in paraffin for optimal BRDU label preservation. I would recommend a slow embedding method for these bones - you can check our article in the JOH, June 2004, Vol. 27(2) pp119-130 which describes our slow embedding method for rat bones which can be adapted for rabbit bones. You can contact me for suggestions and with questions if you would like to. ___ 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] Reusing specimen containers
I have no idea what CAP or JCAHO says about this but it's a very bad idea. Cross contamination happens. If you reuse containers -- and I don't care what precautions you take -- you will rarely or occasionally fail to adequately clean a container and you will end up mixing patient tissues. The cost of a specimen container is a minute fraction of the cost of processing a specimen. Ask yourself how much money you're really saving. You're scrapping for pennies. Or is this some sort of green initiative? In any case, it's asking for problems Daniel Schneider On Tue, Apr 21, 2009 at 11:43 AM, Kelly Colpitts kelly_colpi...@hotmail.com wrote: Hi Histoland! I'm just wondering what folks out there are doing about specimen containers. Is anyone cleaning them out and reusing them? Is there any CAP or Joint Commission regulations that say that all specimen containers can only be used once or can you reuse them as long as they have been thoroughly cleaned? Thanks for you all your input, Kelly _ Windows Live™ SkyDrive™: Get 25 GB of free online storage. http://windowslive.com/online/skydrive?ocid=TXT_TAGLM_WL_skydrive_042009___ 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
