Re: [Histonet] Re: slide printers
We are currently demoing the Fisher Slidemate slide printer and so far we love it. Works well at individual stations but probably not for large batch printing. Mark A Burton HTL ASCP Mass. General Hospital Boston, MA Quoting Lizbeth Kelly lizbeth_ke...@hgsi.com: Hi, Can someone in histoland share names of slide printers that are satisfied with? I prefer no etchers. Thanks, Lizbeth Kelly Human Genome Sciences Rockville, Maryland ___ 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] sharpen permanent knife blades
Please help:-). I am looking for information on anyone who sharpens permanent knife blades. Thanks Marcy CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for use by the designated recipients named above. They are intended solely for these recipients. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Calvert Memorial Hospital immediately by telephone at (410) 535-8282 and destroy all copies of this communication and any attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] sharpen permanent knife blades
C.L. Sturkey (www.sturkey.com) in Lebanon, Pennsylvania, sharpens permanent microtome knives. They also have two grades of disposable blades. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of MARCELLYN STONE Sent: Tuesday, December 22, 2009 9:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sharpen permanent knife blades Please help:-). I am looking for information on anyone who sharpens permanent knife blades. Thanks Marcy CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for use by the designated recipients named above. They are intended solely for these recipients. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Calvert Memorial Hospital immediately by telephone at (410) 535-8282 and destroy all copies of this communication and any attachments. ___ 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] sharpen permanent knife blades
I will also add that DDK and Dorn and Hart Microedge also sharpen knives. One should also check the pricing as they all vary. Jack Sent from my iPhone On Dec 22, 2009, at 8:07 AM, Smith, Allen asm...@mail.barry.edu wrote: C.L. Sturkey (www.sturkey.com) in Lebanon, Pennsylvania, sharpens permanent microtome knives. They also have two grades of disposable blades. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet- boun...@lists.utsouthwestern.edu] On Behalf Of MARCELLYN STONE Sent: Tuesday, December 22, 2009 9:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sharpen permanent knife blades Please help:-). I am looking for information on anyone who sharpens permanent knife blades. Thanks Marcy CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for use by the designated recipients named above. They are intended solely for these recipients. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Calvert Memorial Hospital immediately by telephone at (410) 535-8282 and destroy all copies of this communication and any attachments. ___ 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] Trichrome stain on MMA sections
You can do HE and VVG quite nicely on this type of MMA embedded tissue. Maybe you could also try a Sanderson's (methylene blue) with the Van Gieson (acid fuchsin w/ picric acid) counterstain??? On Dec 21, 2009, at 11:51 PM, Randall Carpenter rjc...@usiwireless.com wrote: Dear Histonet, I was wondering what the best Trichrome stain might be for sawn and ground sections of large stent/artery in methylmethacrylate. I am also looking to stain elastic fibers. Any suggestions? Thanks. Randy Carpenter ___ 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] sharpen permanent knife blades
Different companies have varied angles on how a knife is sharpened for their equipment. A knife sharpened by one company will generally be somewhat different in the angle of the edge than another when you begin cutting. Some may have a flatter edge or a fatter edge at the top cutting portion. These are not always easy to see and can cause issues with sectioning although sharpness is the same or very close. Using several different companies can lead to problems in adjusting the knife angle for sectioning from one to another. I would select one company and stay with it to avoid these issues. I found a company I preferred and then stuck with them rather than fight the problems over a few dollars. Pam Marcum - Original Message - From: Jack Ratliff ratliffj...@hotmail.com To: Allen Smith asm...@mail.barry.edu Cc: Histonet@lists.utsouthwestern.edu, mst...@cmhlink.org Sent: Tuesday, December 22, 2009 9:06:03 AM GMT -06:00 US/Canada Central Subject: Re: [Histonet] sharpen permanent knife blades I will also add that DDK and Dorn and Hart Microedge also sharpen knives. One should also check the pricing as they all vary. Jack Sent from my iPhone On Dec 22, 2009, at 8:07 AM, Smith, Allen asm...@mail.barry.edu wrote: C.L. Sturkey (www.sturkey.com) in Lebanon, Pennsylvania, sharpens permanent microtome knives. They also have two grades of disposable blades. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet- boun...@lists.utsouthwestern.edu] On Behalf Of MARCELLYN STONE Sent: Tuesday, December 22, 2009 9:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sharpen permanent knife blades Please help:-). I am looking for information on anyone who sharpens permanent knife blades. Thanks Marcy CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for use by the designated recipients named above. They are intended solely for these recipients. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Calvert Memorial Hospital immediately by telephone at (410) 535-8282 and destroy all copies of this communication and any attachments. ___ 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
RE: [Histonet] sharpen permanent knife blades
I agree with most of what you have stated and espcially with sticking with one company once you get things moving in the right direction. However, if one truly knows how to properly set and/or adjust cutting angles at the microtome, I would definely keep cost at a high consideration if quality is uniform across vendors. Of course, everyone also has various demands as related to workload volume and study turnaround time. These variables should also be considered and again, especially if product quality is not compromised. Jack Ratliff Date: Tue, 22 Dec 2009 15:36:43 + From: mucra...@comcast.net To: ratliffj...@hotmail.com CC: Histonet@lists.utsouthwestern.edu; mst...@cmhlink.org; asm...@mail.barry.edu Subject: Re: [Histonet] sharpen permanent knife blades Different companies have varied angles on how a knife is sharpened for their equipment. A knife sharpened by one company will generally be somewhat different in the angle of the edge than another when you begin cutting. Some may have a flatter edge or a fatter edge at the top cutting portion. These are not always easy to see and can cause issues with sectioning although sharpness is the same or very close. Using several different companies can lead to problems in adjusting the knife angle for sectioning from one to another. I would select one company and stay with it to avoid these issues. I found a company I preferred and then stuck with them rather than fight the problems over a few dollars. Pam Marcum - Original Message - From: Jack Ratliff ratliffj...@hotmail.com To: Allen Smith asm...@mail.barry.edu Cc: Histonet@lists.utsouthwestern.edu, mst...@cmhlink.org Sent: Tuesday, December 22, 2009 9:06:03 AM GMT -06:00 US/Canada Central Subject: Re: [Histonet] sharpen permanent knife blades I will also add that DDK and Dorn and Hart Microedge also sharpen knives. One should also check the pricing as they all vary. Jack Sent from my iPhone On Dec 22, 2009, at 8:07 AM, Smith, Allen asm...@mail.barry.edu wrote: C.L. Sturkey (www.sturkey.com) in Lebanon, Pennsylvania, sharpens permanent microtome knives. They also have two grades of disposable blades. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Podiatric Medicine -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet- boun...@lists.utsouthwestern.edu] On Behalf Of MARCELLYN STONE Sent: Tuesday, December 22, 2009 9:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sharpen permanent knife blades Please help:-). I am looking for information on anyone who sharpens permanent knife blades. Thanks Marcy CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for use by the designated recipients named above. They are intended solely for these recipients. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Calvert Memorial Hospital immediately by telephone at (410) 535-8282 and destroy all copies of this communication and any attachments. ___ 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
RE: [Histonet] IHC QC slide management
We have a column on the quality control sheet we give the pathologist with the slides. The pathologist marks the column for + patient slide. Joyce Cline, Technical Specialist Hagerstown Medical Laboratory 301-665-4980 fax 301-665-4941 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cheryl [tkngfl...@yahoo.com] Sent: Monday, December 21, 2009 7:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC QC slide management Hi Guys- You're all batting 500 so here's another query: We run a lot of Immunos, not a huge variety but we need to manage how we track patients that are positive and can be used for controls when needed. Does anyone have a simple spreadsheet or concept I could put into a document to make this easy for the people who rotate through this bench to manage well? LOVE labs at Christmas--we're running on chocolate!!! Cheryl Kerry, HT(ASCP) ___ 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] Antigen retrieval and myosin heavy chain question
Dear Histonetters, I was wondering if anyone out there has used antigen retrieval methods to facilitate immuno-staining of formalin-fixed skeletal muscle fibers? I am using ALD-58 + ABC Elite Mouse kit to stain slow-tonic fibers in fresh amphibian muscles, but need to take a look at the fiber distribution of some rare museum specimens. Any suggestions are welcome. Thanks! -Mike -- Mike Jorgensen PhD Candidate Department of Biological Sciences Ohio University Athens, Ohio 45701 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] BK Virus Antibody
We used to purchase our BK virus antibody from a company called Chemicon. Apparently they were bought out by Millipore and the antibody is not available from them. I need to get the antibody from a new source. Any ideas? James Vickroy BS, HT(ASCP) Surgical and Autopsy Pathology Technical Supervisor Memorial Medical Center 217-788-4046 This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Trichrome stain for MMA and Sandersons Rapid Bone Stain chemical makeup (long reply with protocol)
Jack Ratiff wrote: You can do HE and VVG quite nicely on this type of MMA embedded tissue. Maybe you could also try a Sanderson's (methylene blue) with the Van Gieson (acid fuchsin w/ picric acid) counterstain??? On Dec 21, 2009, at 11:51 PM, Randall Carpenter rjcarp http://lists.utsouthwestern.edu/mailman/listinfo/histonet @t usiwireless.com wrote: http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dear Histonet, http://lists.utsouthwestern.edu/mailman/listinfo/histonet http://lists.utsouthwestern.edu/mailman/listinfo/histonet I was wondering what the best Trichrome stain might be for sawn and http://lists.utsouthwestern.edu/mailman/listinfo/histonet ground sections of large stent/artery in methylmethacrylate. I am http://lists.utsouthwestern.edu/mailman/listinfo/histonet also looking to stain elastic fibers. Any suggestions? Thanks. http://lists.utsouthwestern.edu/mailman/listinfo/histonet http://lists.utsouthwestern.edu/mailman/listinfo/histonet Randy Carpenter http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Several years ago I posted a discussion of Sanderson's bone stain is the same as Stevenels blue developed for bone work by Maniatopoulos C, Rodriguez A, Deporter DA, Melcher AH: An improved method for preparing histological sections of metallic implants. Internat J Oral Maxillofacial Implants 1(1):31, 1987. The stain was originally developed as a parasite stain. Sanderson figured out an easier way to make up the staining solution than the original recipe, and then it was marketed by Surgipath with trademark. Her recipe is proprietary and stains with the same results as the Maniatopoulos method. We did the comparison in our lab and had identical results. Because the Stevenel's is such a royal pain to make in the lab, I suggest buying the Sanderson Bone stain. It is money well spent to avoid a long day of stain preparation. The chemistry of making this stain is interesting in that potassium permangante oxidizes methylene blue, forming a thick gooey ppt that takes a great deal of stirring and heating to get things into solution. The pH is very alkaline, somewhere in 9 or higher range and when using this stain, further heating of the solution the methylene blue will continue to oxidize and the pH continues to increase. We found our homemade Stevenels needed to be topped off frequently, and also filtered since a black ppt keeps forming. Conn's Biological stains Lillie, RD, revised by Stotz EH and Emmel,VM: H J Conns Biological Stains. pp 423-424, Ninth ed., Williams and Wilkins Co, Baltimore MD 1977 has explanation of what happens to the methylene blue when oxidized by KMNO4. The by- products of the methylene blue oxidation are toluidine blue, methylene violet, thionin, Azure A and other Azures along with residual methylene blue left in the solution. Some dyes are often found in formulations/recipes for PMMA embedded bone sections e.g. toluidine blue. Staining results are Osteoid - blue to intense blue-green; Muscle, connective tissue - blue to blue-green; Cartilage - blue and/or shades of violet to purple; Calcified cartilage - medium to dark purple Calcified bone is stained by acid fuchsin, and light green can also be used. Basic Fuchsin is also a counterstain Since PMMA is very hydrophobic, only low molecular weight dyes penetrate the plastic sufficiently in order to stain all the the described components. Acid fuchsin also has a low molecular weight. RW Horobin has a wonderful publication on the effects of staining on plastics. I am not putting my finger on the reference, but this paper alone is an education on how dyes work on plastic embedded tissues, including PMMA and GMA. Some important factors that affect staining of PMMA embedded bone are Low molecular weight dyes Temperature - must be heated pH - alkaline is important for good staining. These factors also affect how hydrophilic plastics for EM and other methacrylate embedded tissues stain. It is well established that toluidine blue, in sodium borate at pH 11 is a stain of choice for EM embedded tissues, and requires heating. Hopefully this will help in understanding the mechanism. I suggest accessing the publications for further information. http://lists.utsouthwestern.edu/mailman/listinfo/histonet Personally, I have preferred MacNeals Tetrachrome over Sandersons/Stevenels for undecalcified bone in MMA, ground sections but the surface must be etched with 0.7% formic acid and also alcohol etching of MMA, rinsed well, dried then immersed into the MacNeals. Do not buy MacNeals as a commercial preparation, it does not work as well as an in house preparation. Here is the protocol for MacNeals with a basic fuchsin counterstain. MacNeals can also be combined with Toluidine blue for a brilliant, well stained bone section in MMA.
[Histonet] Happy Holidays
To all my friends and colleagues on Histonet I wish you a very Happy Holiday season filled with family and friends Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Directly conjugated secondaries versus biotinylated secondaries
Greetings, Hopefully all of you are enjoying a great time with your friends and family this week rather than working like me. Here is my problem. I have an antibody (anti-panendothelial antigen, i.e. MECA32 from BD http://www.bdbiosciences.com/ptProduct.jsp?prodId=19506) and I'm trying to get it to work on 4% PFA fixed, decalcified, paraffin embedded sections. I previously titered it to 2 ug / mL with a biotinylated anti-rat F(ab')2 (1 ug / mL) and then a strepavidin Dylight 594 (1 ug / mL) and got beautiful, strong staining. Those of you who have been helping me from my IHC infancy would be proud. However, I really need it to be a directly conjugated secondary because I want to co-stain with something else that only works with a biotinylated secondary followed by tyramide amplification. So I retitered it from 5 ug / mL and down and came in with a Dylight-649 conjugated anti-rat (1 ug / mL) secondary and I saw nothing (Dylight 649, which is a Cy5 replacement, is too far red to see without the aid of a camera--although the guy who helped me use the microscope says there's one woman on campus who has infrared vision). I should note that another group has reported getting this to work with a biotinylated primary and adding an fluorophore conjugated avidin. I can think of a couple of possibilities here 1) The antibody needs that extra edge of a biotinylation to get sufficient signal (has anyone ever seen this problem which couldn't be solved by titrating something?) 2) I need to increase the primary titer ever more 3) I need to increase the secondary titer (what primary titer should I use if I do this?) 4) Dylight 649 is just too dim to see anything and if I switched to another fluor, it would work. I have no experience with this fluor, as our floor scope doesn't have the filters for it. 5) This is all a fluke and I screwed something up Any suggestions on how to troubleshoot this are welcome. Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
