[Histonet] Microtomist's Vade-Mecum on the Web
Arthur Bolles Lee's The Microtomist's Vade-Mecum, 7th edition (1913) is available online on Google Books, along with some other editions: http://books.google.com/books?id=4m5MMAAJprintsec=frontcoverdq=arthur+bolles+leeei=BpJhS96xKJTIzASV9vnNBQcd=1#v=onepageq=f=false This marvelous old histology recipe book was a great rarity before the Web, though it's actually now available from Amazon. It took me several years to find a copy in 1976 - found one in a consignment of old books discarded from the library of a long-defunct college. A note about the title: Vade-Mecum (pronounced something like voddy-make-um) - literally come with me (think of Quo Vadis) is an old word for a practical handbook of something. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Stains for Macrophages for Laser CaptureMicrodissection purpose
Delphine, I think John is right about identifying the cells by shape. I do LCM, and I perform a rapid hematoxylin stain just for orientation purposes. The total time from fixation to air drying is 7 minutes. Any more time than that, and the RNA is too degraded to get good results from downstream applications. You could substitute toluidine blue for the hematoxylin in this case. Are you really looking for a (no greater than) 30 minute (aqueous) stain? Seems way too long for good quality and quantity RNA. I would be interested in hearing what you usually do for staining for LCM purposes and what your RNA results have been. Karen Karen Percival, BS, HT Research Scientist II Pfizer Research DSRD 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com John Kiernan jkier...@uwo.ca 1/27/2010 10:41 PM How about phase contrast optics and identifying the cells by their shapes? Non-specific addition of colour to a section of unfixed tissue probably will not show the cells any more clearly. I have taken the liberty of including a colleague, Dr Paul Walton, in this reply. He does laser capture microdissection and may have something to suggest. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: delphine eberle eberledelph...@hotmail.com Date: Wednesday, January 27, 2010 16:27 Subject: Stains for Macrophages for Laser Capture Microdissection purpose To: jkier...@uwo.ca, sharon.will...@bms.com Cc: histonet@lists.utsouthwestern.edu Hi, I have another question following Macrophage staining. I am setting up a Laser Capture Microdissection protocol to dissect macrophages from atherosclerosis lesions (non fixed frozen sections) and extract RNA for gene expression purpose. I am looking at a quick staining (less than 30min otherwise RNA is degradated) for macrophages in that context? Any suggestions? Thanks a lot, Delphine Delphine Eberlé PhD UCSF Department of Vascular Surgery eberledelph...@hotmail.com From: jkier...@uwo.ca To: sharon.will...@bms.com Date: Wed, 27 Jan 2010 00:22:53 -0500 Subject: Re: [Histonet] Histology Special Stains for Macrophages CC: histonet@lists.utsouthwestern.edu None of the methods mentioned in the enquiry are stains for macrophages. Research workers who never took Histology 101 often stain cells of the monocyte/macrophage lineage immunohistochemically (IHC), using very expensive primary antibodies and fairly expensive kits to amplify and detect the binding sites. IHC is necessary if you must find every macrophage, including a tissue's recent monocyte immigrants that haven't yet done any work. Macrophages that have been busily eating blood are full of brown granules that don't need any staining. Ordinary people recognize macrophages by their appearance in sections stained with a well done HE or with one of the many Romanowsky-Giemsa blood stains. With the latter it's important to get the pH right - much lower for sections of formaldehyde-fixed objects (pH4 to 5) than for methanol-fixed films or smears (traditionally 6.8). It's all very well explained in RD Lillie GM Fullmer's Histopathologic Technic... (4th ed 1976, pp.193-197; the 3rd edition, 1965, is OK too) and also in more recent textbooks including those by Bancroft Gamble, Geoffrey Brown, Freida Carson and, of course Yours truly. John Kiernan Anatomy, UWO London, Canada http://publish.uwo.ca/~jkiernan/bookfind.htm = = = - Original Message - From: Willman, Sharon sharon.will...@bms.com Date: Tuesday, January 26, 2010 12:12 Subject: [Histonet] Histology Special Stains for Macrophages To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Hi, We are needing to do a special stain for macrophages. What is the most common stain for that? Does anyone do a Sudan Black, Alcian Blue or Van Gieson for macrophages? Any information would be appreciated. Thanks in advance. Sharon This message (including any attachments) may contain confidential, proprietary, privileged and/or private information. The information is intended to be for the use of the individual or entity designated above. If you are not the intended recipient of this message, please notify the sender immediately, and delete the message and any attachments. Any disclosure, reproduction, distribution or other use of this message or any attachments by an individual or entity other than the intended recipient is prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu
[Histonet] Re: [IHCRG] Hamamatsu Nanozoomer
I would like to thank everyone that responded to my questions about these scanners. These answers have been very valuable to my boss and I and will be taken into account when we decide on which system to purchase! Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: James Watson jwat...@gnf.org To: Kim Merriam kmerriam2...@yahoo.com Cc: joel.duckwo...@olympus.com joel.duckwo...@olympus.com Sent: Thu, January 28, 2010 9:15:52 AM Subject: RE: [IHCRG] Hamamatsu Nanozoomer Kim, I use the nanozoomer a lot, we have scanned about 45,000 images in 3 years. I bought it because it does both fluorescence and transmitted light very well. The aperio scanner is good and the software can view both types of scans. The Nanzoomer software has improved greatly in the past 3 years thanks to Olympus and the service has been excellent. I will send you some images and more information when I get to work. I have CC’ed my sales rep, he will make sure someone gets in touch with you today. The cost is about the same as a single instrument from Aperio (unless aperio changed their pricing). Jamie From:Kim Merriam [mailto:kmerriam2...@yahoo.com] Sent: Thursday, January 28, 2010 4:44 AM To: James Watson Subject: Re: [IHCRG] Hamamatsu Nanozoomer Thanks Jamie. When I was at Novartis/NIBRI, I used the Aperio quite a bit (I am at Amgen now, but I remember meeting you a while back at one of the NSH meetings). Anyway, the other Amgen sites use the Aperio, but we are intrigued by the possibility of buying one machine that can do light and IF imaging, also the Z-stack feature is a nice ad-on (although I am not sure how much we would really use it). Based on the information I have gathered, I would be able to pull the images taken from the Nanozoomer and put them on our Aperio server for cross-site conferences. We were actually ready to purchase the Aperio next week, but then my boss found out about the Nanozoomer, which seems to be a better instrument, but I have no idea how much it costs, I cant get anyone from Hamamatsu or Olympus to call me back. Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From:James Watson jwat...@gnf.org To: kmerriam2...@yahoo.com kmerriam2...@yahoo.com Sent: Thu, January 28, 2010 1:08:16 AM Subject: FW: [IHCRG] Hamamatsu Nanozoomer Kim, Contact me, I have a Nanozoomer and also know a lot about the Aperio since their headquarters is near here and I have been to their office many times and they have been here. Jamie James Watson HT ASCP Facilities Manager of Histology GNF Genomics Institute of the Novartis Research Foundation Tel 858-332-4647 Fax 858-812-1915 jwat...@gnf.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Wednesday, January 27, 2010 9:09 AM To: kmerriam2...@yahoo.com; Histonet; ih...@googlegroups.com Subject: [Histonet] RE: [IHCRG] Hamamatsu Nanozoomer Kim We have an Aperio Scanner. I'm not quite sure what you are asking but this is what I think from your e-mail. Aperio scans create .svs files these files are considered a modified tiff file. When aperio scans the file is placed into the spectrum database - from there you can create case or project files, etc. These images can be viewed via imagescope. There is no issue with placing other images that have not been generated on the scanscope into the spectrum database, you can upload any image into the spectrum database we have done this in the past. These images can also be viewed with imagescope we have only uploaded single magnification tiff files into the aperio spectrum database. The Hamamatsu Nanozoomer I believe also comes with a database and some viewing sofware. Are you asking if you can view the HN images in the spectrum database? To be honest I'm not sure if you can view the images completed with image scope I would ask your sales rep. I'm sure you are aware that both of these scanners essentially create virtual slides. Not just an image of a slide at one magnification. The viewing software allows you to view the images at different magnifications. I'm not that familar with Visiomorph its the Olympus product and doesn't Olympus sell the HN scanner? I would also think that that the database that comes with the HN scanner should be able to accept other images and should have some conferencing capabilities, I know that the aperio does. I'm not sure if I answered your questions, but I would ask the sales reps from both aperio and Olympus about compatability. With repects to analysis I think that the Visiomorph software will be able to analyze a image generated from the scanscope. If you are working with an HN scanner and the visiomorph software I'm assuming that you will be able to perform batch analysis on selected images from the olympus database. You
[Histonet] Re: Diff-Quik
Dear Colleagues, At the risk of beating a dead horse, I would like to thank Bryan Llewellyn for so eloquently and succinctly echoing my own thoughts on the matter that Dr. Richmond commented on which has drawn criticism. For those who may have missed it, I'll include it below my own meager response. Teri Johnson, HT(ASCP)QIHC Managing Director, Histology Facility Stowers Institute for Medical Research Kansas City, MO Date: Wed, 27 Jan 2010 13:57:16 -0800 From: Bryan Llewellyn llewl...@shaw.ca Subject: Re: [Histonet] Re: Diff-Quik To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 26938660d7e848faa055bcb0a76df...@bryanpc Content-Type: text/plain; format=flowed; charset=iso-8859-1; reply-type=original I have worked in several laboratories, both in Canada and England. I trained there and was taught by technologists, not pathologists, to check the staining of all my slides, which I have always done. I finds that even when asked, most pathologists do not return poorly stained slides, yet how can we know of deficiencies if they do not? I worked in several histology labs over my career and slides were checked in all but two. I was able to introduce it in one of those, but not in the other due to technologist resistance (we've always done it that way). It was the technologists who resisted it. I don't see why Dr. Richmond needs to apologise. He merely drew attention to his work experiences in numerous histology labs and decried what is, in his experience, a common practice. In other words, he spoke about real, actual events. Why does being truthful require an apology? Are our egos so fragile that we can't take valid criticism as a profession nor as individuals? He certainly did not accuse those who got upset of failing to check slides, so why take umbrage? Noting and decrying poor histological practices should be a concern for everyone. Surely we are concerned about service to patients not about our egos. Bryan Llewellyn ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fw: [IHCRG] Dianova rat anti-mouse CD31
Hi - someone emailed me about my methodology, but I cant find the email, so here it is: I tried it @ 1:10, 1:25 and 1:50. All seem viable dilutions for the FFPE xenografts. I used BIocare DIVA (EDTA) HIER in decloaking chamber, primary ab for 2 hours @ RT, Biocare Rat-on-Mouse HRP polymer kit (15 minutes each) + DAB. The path labs (for my company) that are in California and Seattle used the same retreival but different detection methods and got similar results. Kim From:ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of Kim Merriam Sent: Wednesday, January 27, 2010 9:23 AM To: Histonet; ih...@googlegroups.com Subject: [IHCRG] Dianova rat anti-mouse CD31 Hi All, I know there was some buzz a while back about this antibody. I have tested it on mouse FFPE xenografts, and I must say, it looks very promising. I compared 2 pieces of the same mouse xenograft; one half of the tumor was fixed in IHC-Zinc (ZInc-Tris) and stained with the BD rat anti-mouse CD31 and the other half of the tumor was fixed in NBF. The NBF-fixed tumor actually looks better! A couple of other folks that are working at the west coast sites of my company have had similar results. We are all very encouraged. I still need to do a bit more testing with some other tissues (and perhaps some additional titrations), but I thought I would pass this along to everyone. Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Kp Marker Plus Slide Pen
Good morning fellow Histotechs, Does anyone know of a supplier for the KP Maker Plus slide pens? Any information on this would be very helpful thanks as I can not seem to locate a vendor. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Kp Marker Plus Slide Pen
Mercedes Medical -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of brent hart Sent: Thursday, January 28, 2010 10:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Kp Marker Plus Slide Pen Good morning fellow Histotechs, Does anyone know of a supplier for the KP Maker Plus slide pens? Any information on this would be very helpful thanks as I can not seem to locate a vendor. ___ 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] Fwd: Re: Stains for Macrophages for Laser Capture Microdissection purpose
- Original Message -
From: Christopher Pin c...@uwo.ca
Date: Thursday, January 28, 2010 9:22
Subject: Re: Stains for Macrophages for Laser Capture Microdissection purpose
To: Martin Sandig martin.san...@schulich.uwo.ca, John Kiernan
jkier...@uwo.ca, Paul Walton pwal...@uwo.ca
Hi all,
Thanks for including me in these discussions. I am no expert on macrophage
histology but I know something about laser capture microdissection. The key
to this method for RNA isolation is not so much in the timing but rather in
the solutions complete lack of RNAse. We have worked with pancreatic tissue
and been able to dissect of cells for RNA isolation. Likely one approach
would be to do HE for identification. Before you went to the trouble of
doing LCM, you could take some unimportant tissue, scrape the tissue off the
slide after fixation and then isolate RNA just to verify the integrity of the
RNA following staining.
The other thing that you can do is fix the slides as if you are doing in situ
hybridization before you do any histology.
I know this isn’t that helpful but from my experience it is not the time but
rather the quality of the solutions that makes all the difference.
Chris
On 1/28/10 9:01 AM, Martin Sandig martin.san...@schulich.uwo.ca wrote:
Hi:
I do not do laser capture at all.
For identification purposes, perhaps it would be possible to load the
macrophages with an injected dye (they eat that stuff depending on the tissue
under investigation) and then capture them. In atherosclerotic lesions they
are loaded with lipids (foam cells) and perhaps with an excellent microscope
they may be visible before capture.
Chris Pin (c...@uwo.ca) may have some experience with laser micro-dissection.
Cheers,
Martin
Martin Sandig, PhD
Associate Professor
Associate Chair, Undergraduate Affairs
Department of Anatomy and Cell Biology
Schulich School of Medicine and Dentistry
University of Western Ontario
Dental Sciences Building, Room 00075
Phone: 519 661 2111 86815
Fax: 519 850 25662
http://www.uwo.ca/anatomy/department/sandigm/msandig.html
Paul Walton pwal...@uwo.ca 28/01/2010 8:24 am
John,
I have forwarded this message on to Martin, who does laser capture. Alas, I
am the microinjection fellow.
Paul
Begin forwarded message:
From: John Kiernan jkier...@uwo.ca
Date: January 27, 2010 10:41:15 PM EST (CA)
To: delphine eberle eberledelph...@hotmail.com
Cc: sharon.will...@bms.com, histonet@lists.utsouthwestern.edu, pwal...@uwo.ca
Subject: Re: Stains for Macrophages for Laser Capture Microdissection purpose
How about phase contrast optics and identifying the cells by their shapes?
Non-specific addition of colour to a section of unfixed tissue probably will
not show the cells any more clearly. I have taken the liberty of including a
colleague, Dr Paul Walton, in this reply. He does laser capture
microdissection and may have something to suggest.
John Kiernan
Anatomy, UWO
London, Canada
= = =
- Original Message -
From: delphine eberle eberledelph...@hotmail.com
Date: Wednesday, January 27, 2010 16:27
Subject: Stains for Macrophages for Laser Capture Microdissection purpose
To: jkier...@uwo.ca, sharon.will...@bms.com
Cc: histonet@lists.utsouthwestern.edu
Hi,
I have another question following Macrophage staining.
I am setting up a Laser Capture Microdissection protocol to dissect
macrophages from atherosclerosis lesions (non fixed frozen sections) and
extract RNA for gene expression purpose.
I am looking at a quick staining (less than 30min otherwise RNA is
degradated) for macrophages in that context? Any suggestions?
Thanks a lot,
Delphine
Delphine Eberlé PhD
UCSF Department of Vascular Surgery
eberledelph...@hotmail.com java_script:main.compose('new',
't=eberledelph...@hotmail.com')
From: jkier...@uwo.ca
To: sharon.will...@bms.com
Date: Wed, 27 Jan 2010 00:22:53 -0500
Subject: Re: [Histonet] Histology Special Stains for Macrophages
CC: histonet@lists.utsouthwestern.edu
None of the methods mentioned in the enquiry are stains for macrophages.
Research workers who never took Histology 101 often stain cells of the
monocyte/macrophage lineage immunohistochemically (IHC), using very
expensive primary antibodies and fairly expensive kits to amplify and
detect the binding sites. IHC is necessary if you must find every
macrophage, including a tissue's recent monocyte immigrants that haven't
yet done any work. Macrophages that have been busily eating blood are
full of brown granules that don't need any staining.
Ordinary people recognize macrophages by their appearance in sections
stained with a well done HE or with one of the many Romanowsky-Giemsa
blood stains. With the latter it's important to get the pH right - much
lower for sections of formaldehyde-fixed objects (pH4 to 5) than for
RE: [Histonet] Kp Marker Plus Slide Pen
Yeah, that's it - that's where we got ours from. Plus the Mercedes Medical rep called to remind me after seeing my post, so he does monitor the Histonet... ;-) --On Thursday, January 28, 2010 10:39 AM -0500 Blazek, Linda lbla...@digestivespecialists.com wrote: Mercedes Medical -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of brent hart Sent: Thursday, January 28, 2010 10:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Kp Marker Plus Slide Pen Good morning fellow Histotechs, Does anyone know of a supplier for the KP Maker Plus slide pens? Any information on this would be very helpful thanks as I can not seem to locate a vendor. ___ 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 Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) 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
RE: [HISTONET] embedding cell cultures
Hi Nick, If you can, grow the cells on thermanox cover slips (NUNC). Then process them as normal for embedding in Eponate 12. I use acetonitrile in place of the propylene oxide to avoid damaging the coverslip. When doing the final embedding and polymerization, invert the coverslip over a drop of resin on a square of ACLAR sheeting. This forms a sandwich of your cell layer. After polymerization you separte the resin layer from the ACLAR and the coverslip (very easy! No sawing!) and re-embed one or more layers on their side in a resin block. Now, section away. You will get beautiful sections of your cells at a nice 90 degree angle. The best part of doing it this way is that you can put the embedded cell layer in the light microscope and select any area of interest before re-embedding and sectioning. Good luck! Jo Dee ~~Jo Dee Fish~~ Senior Research Technologist The J. David Gladstone Institutes Co-manager Histology and Microscopy Core Telephone: (415) 734-2567 Fax: (415) 355-0824 E-mail: jf...@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 Nicholas David Evans wrote: Dear all, I was hoping someone might be able to offer me some advice on embedding and sectioning cell cultures. In short we are growing cells which form 3D dome-like structures on tissue culture plastic. Does anyone have any experience or advice to offer on embedding the cultures in situ before sectioning? I have seen various methods in the literature, which often use Epon to embed the material followed by sawing away the plastic, but if anyone can offer some tips on other possible (easier) ways of doing it, or can refer me to some useful literature, I'd be very grateful. I would like to have simple 10um sections at 90 degrees to the substrate, which I can use for IHC. Best wishes Nick ___ 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] Kp Marker Plus Slide Pen
They last so long! That's why it's easy to forget where they came from! -Original Message- From: Maria Katleba [mailto:maria.katl...@stjoe.org] Sent: Thursday, January 28, 2010 11:56 AM To: Merced M Leiker; Blazek, Linda; 'brent hart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Kp Marker Plus Slide Pen These are the BEST pens I have ever bought for Histology! They last way longer plus the writing is very black and the colour stays... even when you reprocess a block Yes... I mark my kids school supplies with them too! Mercedes Medical 1-800-331-2716 www.MercedesMedical.com These are all I ever buy anymore... no one comes close. Maria Katleba HT (ASCP) MS Queen of the Valley Medical Center Napa CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Merced M Leiker Sent: Thursday, January 28, 2010 8:14 AM To: Blazek, Linda; 'brent hart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Kp Marker Plus Slide Pen Yeah, that's it - that's where we got ours from. Plus the Mercedes Medical rep called to remind me after seeing my post, so he does monitor the Histonet... ;-) --On Thursday, January 28, 2010 10:39 AM -0500 Blazek, Linda lbla...@digestivespecialists.com wrote: Mercedes Medical -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of brent hart Sent: Thursday, January 28, 2010 10:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Kp Marker Plus Slide Pen Good morning fellow Histotechs, Does anyone know of a supplier for the KP Maker Plus slide pens? Any information on this would be very helpful thanks as I can not seem to locate a vendor. ___ 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 Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) 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 Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Kp Marker Plus Slide Pen
These are the BEST pens I have ever bought for Histology! They last way longer plus the writing is very black and the colour stays... even when you reprocess a block Yes... I mark my kids school supplies with them too! Mercedes Medical 1-800-331-2716 www.MercedesMedical.com These are all I ever buy anymore... no one comes close. Maria Katleba HT (ASCP) MS Queen of the Valley Medical Center Napa CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Merced M Leiker Sent: Thursday, January 28, 2010 8:14 AM To: Blazek, Linda; 'brent hart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Kp Marker Plus Slide Pen Yeah, that's it - that's where we got ours from. Plus the Mercedes Medical rep called to remind me after seeing my post, so he does monitor the Histonet... ;-) --On Thursday, January 28, 2010 10:39 AM -0500 Blazek, Linda lbla...@digestivespecialists.com wrote: Mercedes Medical -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of brent hart Sent: Thursday, January 28, 2010 10:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Kp Marker Plus Slide Pen Good morning fellow Histotechs, Does anyone know of a supplier for the KP Maker Plus slide pens? Any information on this would be very helpful thanks as I can not seem to locate a vendor. ___ 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 Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) 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 Notice from St. Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] I am no longer receiving messages from the histonet
Kaye Ryan North Florida Dermatology Associates Histology Lab Supervisor (904) 398-0547 Ext. 2004 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: [IHCRG] Dianova rat anti-mouse CD31
Thank you Kim, that does sound very promising, I still have not gotten
around to trying my sample of this, your work will be very helpful when I
do.
Best regards,
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
This email is confidential and intended solely for the use of the Person(s)
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From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
James Watson
Sent: Thursday, January 28, 2010 8:19 AM
To: 'kmerriam2...@yahoo.com'; Histonet; ih...@googlegroups.com
Subject: RE: [IHCRG] Dianova rat anti-mouse CD31
I just got some of this antibody in an will be setting it up on the Ventana
Discovery Thank you for the information on your protocol. I will let you
know how it goes.
Jamie
James Watson HT ASCP
Facilities Manager of Histology
GNF Genomics Institute of the Novartis Research Foundation
Tel858-332-4647
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From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
Kim Merriam
Sent: Thursday, January 28, 2010 6:46 AM
To: Histonet; ih...@googlegroups.com
Subject: Fw: [IHCRG] Dianova rat anti-mouse CD31
Hi - someone emailed me about my methodology, but I cant find the email, so
here it is:
I tried it @ 1:10, 1:25 and 1:50. All seem viable dilutions for the FFPE
xenografts. I used BIocare DIVA (EDTA) HIER in decloaking chamber, primary
ab for 2 hours @ RT, Biocare Rat-on-Mouse HRP polymer kit (15 minutes each)
+ DAB. The path labs (for my company) that are in California and Seattle
used the same retreival but different detection methods and got similar
results.
Kim
From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
Kim Merriam
Sent: Wednesday, January 27, 2010 9:23 AM
To: Histonet; ih...@googlegroups.com
Subject: [IHCRG] Dianova rat anti-mouse CD31
Hi All,
I know there was some buzz a while back about this antibody. I have tested
it on mouse FFPE xenografts, and I must say, it looks very promising. I
compared 2 pieces of the same mouse xenograft; one half of the tumor was
fixed in IHC-Zinc (ZInc-Tris) and stained with the BD rat anti-mouse CD31
and the other half of the tumor was fixed in NBF. The NBF-fixed tumor
actually looks better!
A couple of other folks that are working at the west coast sites of my
company have had similar results. We are all very encouraged.
I still need to do a bit more testing with some other tissues (and perhaps
some additional titrations), but I thought I would pass this along to
everyone.
Kim
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
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[Histonet] Ga.Society For Histotechnology Reservation Link
Hi All, I wanted to pass this link from the hotel on to you guys who will be attending the Georgia Society for Histotechnology meeting March 26-28 this year. This goes straight to the hotel and already has our code entered for you to reserve your rooms at the Evergreen Marriott Conference Resort. For the complete program and registration form go to www.histosearch.com/gshhttp://www.histosearch.com/gsh and go to the symposium page. You will find the hotel link there too. See you there. Shirley Powell GSH Secretary Subject: Ga.Society For Histotechnology Reservation Link Simply cut and paste the link below and include with your electronic correspondence to facilitate the reservation process. Your guests will be directed to the property's home page with the code already entered in the appropriate field. All they need to do is enter their arrival date to begin the reservation process. Evergreen Marriott Conference Resort http://www.marriott.com/hotels/travel/atleg?groupCode=gshgshaapp=resvlinkfromDate=3/26/10toDate=3/27/10 [http://www.marriott.com/Images/Brands/MCC/Logos/MCC_logowhitefield_120x60.gif]http://www.marriott.com/hotels/travel/atleg?groupCode=gshgshaapp=resvlinkfromDate=3/26/10toDate=3/27/10 http://www.marriott.com/hotels/travel/atleg?groupCode=gshgshaapp=resvlinkfromDate=3/26/10toDate=3/27/10 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 74, Issue 34
Dear Nick, When I was growing and harvesting 3D cell cultures (spheroids grown in matrigel) we would remove the culture medium, draw up the gel in a needle-less syringe and place in to OCT embedding medium (Tissue Tek) before freezing in Liquid Nitrogen. I would then cut 5-10um sections and perform IHC or immunofluorescence. Hope this helps - but it looks like you have a lot of good suggestions already. Best Regards, Rhonda Rhonda Henshall-Powell, Ph.D. -- Message: 2 Date: Wed, 27 Jan 2010 10:25:42 -0800 (PST) From: Nicholas David Evans ndev...@stanford.edu Dear all, I was hoping someone might be able to offer me some advice on embedding and sectioning cell cultures. In short we are growing cells which form 3D dome-like structures on tissue culture plastic. Does anyone have any experience or advice to offer on embedding the cultures in situ before sectioning? I have seen various methods in the literature, which often use Epon to embed the material followed by sawing away the plastic, but if anyone can offer some tips on other possible (easier) ways of doing it, or can refer me to some useful literature, I'd be very grateful. I would like to have simple 10um sections at 90 degrees to the substrate, which I can use for IHC. Best wishes Nick -- Message: 3 Date: Wed, 27 Jan 2010 15:19:36 -0500 From: Geoff McAuliffe mcaul...@umdnj.edu Hi Nick: You can use the Epon substitutes such as EmBed 812. Fix, osmicate, dehydrate as usual, but omit the proplylene oxide as it will react with the plastic dish. Epon substitutes will mix with ethanol. I used 2:1 then 1:1 then 1:2 ratios of absolute ethanol to Epon with catalyst added with agitation. Then several changes of pure Epon and polymerize. Yes, you will have to saw away the plastic, if you try to section the Epon-plastic combo the two will separate. How much easier do you want it to be? Geoff -- Message: 5 Date: Wed, 27 Jan 2010 15:39:05 -0500 From: Sherwood, Margaret msherw...@partners.org Subject: RE: [HISTONET] embedding cell cultures Nick, I am assuming that your 3D cells only grow on plastic. They make plastic cover slips which, if your cells attach to them and grow, might make the embedding much easier. You would follow the same method as stated, but then you could invert the coverslips on a beem capsule and separate the coverslip from capsule with liquid nitrogen. However, I have never done it with plastic coverslips (only glass), so not sure if they would easily separate from capsule with liquid nitrogen. If anyone else has done so, please weigh in. Peggy -- Message: 6 Date: Wed, 27 Jan 2010 15:44:37 -0500 From: Peggy Bisher mbis...@princeton.edu Subject: Re: [HISTONET] embedding cell cultures I have done just what you are talking about using Aclar. It is a plastic embedding film (purchased from EMS). It works great for us. Margaret E. Bisher Electron Microscopy Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbis...@princeton.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] need help with stain
A researcher wants to measure the length of the intestinal crypts and asked for a suggestion on what type of stain to use. I am thinking of using a phloxine/tartrazine stain and do a double stain with a GMS to show the basement membrane. What is your opinion on this? Do you have other suggestions that would work better? Hope you are warmer than here in SD! Thanks for the help. Margaret Perry South Dakota State University ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] need help with stain
Use mucicarmin stain. The crypt mucus will be evident and their sizes. René J. --- On Thu, 1/28/10, Perry, Margaret margaret.pe...@sdstate.edu wrote: From: Perry, Margaret margaret.pe...@sdstate.edu Subject: [Histonet] need help with stain To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Thursday, January 28, 2010, 4:00 PM A researcher wants to measure the length of the intestinal crypts and asked for a suggestion on what type of stain to use. I am thinking of using a phloxine/tartrazine stain and do a double stain with a GMS to show the basement membrane. What is your opinion on this? Do you have other suggestions that would work better? Hope you are warmer than here in SD! Thanks for the help. Margaret Perry South Dakota State University ___ 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: [SPAM-HC] - [Histonet] Microtomist's Vade-Mecum on the Web - Email found in subject
Samuri. The website www.scienceheritagelimited.com has available 8 books that are reprints of historically important history of microscopy and histotechnology books included is the 1885 edition of Arthur Bolles Lee's The Microtomist's Vade-Mecum. It's amazing how little (relatively) things have changed. The only thing missing in the 1885 edition is the use of formalin as a tissue fixative. This did not come about until 1892 and undoubtedly appears in his later editions. Good reading, jim J.B.McCormick, M.D. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Thursday, January 28, 2010 7:42 AM To: Histonet@lists.utsouthwestern.edu Subject: [SPAM-HC] - [Histonet] Microtomist's Vade-Mecum on the Web - Email found in subject Arthur Bolles Lee's The Microtomist's Vade-Mecum, 7th edition (1913) is available online on Google Books, along with some other editions: http://books.google.com/books?id=4m5MMAAJprintsec=frontcoverdq=arthur+bolles+leeei=BpJhS96xKJTIzASV9vnNBQcd=1#v=onepageq=f=false This marvelous old histology recipe book was a great rarity before the Web, though it's actually now available from Amazon. It took me several years to find a copy in 1976 - found one in a consignment of old books discarded from the library of a long-defunct college. A note about the title: Vade-Mecum (pronounced something like voddy-make-um) - literally come with me (think of Quo Vadis) is an old word for a practical handbook of something. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thomas Crowell is out of the office.
I will be out of the office starting 01/28/2010 and will not return until 02/02/2010. Please contact Kelly Miner at 617-871-5122 if you have any questions regarding clinical trial samples. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Diff-Quik
Please don't take it personally. If we are trying our best to meet our own high expectations then we do not have to worry. I believe that most of us do what Robert expects. This is supported by my experiences at the recent NSH meeting in Birmingham Alabama. I was privileged to meet some of the most dedicated professionals around and believe that our profession is heading in the right direction. Robert's comments should be heeded and pondered. I definitely did. 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 kim.dona...@bhcpns.org Sent: Thursday, 28 January 2010 2:53 AM To: Robert Richmond Cc: histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Diff-Quik I'm going to have to agree with Cheryl on the comment. This may be your experience but I can tell you my techs always look at their stains before they send it on to the Pathologist. It is a requirement that they understand what they are looking at in order to know if it worked. Each of them are also trained to know all tissues microscopically and all stain components microscopically. That is after all the purpose of being a Histologist. I am going out on a limb here and I normally don't, but you are digging yourself in to a rather rude hole to insult so many professional Histologist. Just saying. Kim Donadio Pathology Supervisor Baptist Hospital 1000 W Moreno St. Pensacola FL 32501 Phone (850) 469-7718 Fax (850) 434-4996 Robert Richmond rsrichm...@gmail.com Sent by: histonet-boun...@lists.utsouthwestern.edu 01/26/2010 07:38 PM To histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu cc Subject [Histonet] Re: Diff-Quik I sort of apologize for this ill-natured comment, which long-term readers of Histonet know I've made before. I do locum tenens work, mostly in rather small pathology services - I've worked in perhaps 60 of them in my life. Only rarely do I observe that a histotech ever looks at a slide. I've just acquired a new client with particularly difficult slides. The tech doesn't even have a microscope. The more quality assurance paperwork I have to do, the worse the slides. The lack of feedback from pathologist to technologist is a really widespread and serious problem. Most pathologists are completely unwilling to take the time to do it, and the usage has never established itself. It would be much easier if we had double headed microscopes, which seem to be prohibited in small pathology services. Did Edwards Deming live in vain? Bob Richmond Samurai Pathologist Knoxville TN * On Tue, Jan 26, 2010 at 9:03 AM, Cheri Miller cmil...@physlab.com wrote: Every slide I stain, special stains, IHC or otherwise I check under the scope...I have taught all my techs to do the same, other than batches of HE and then we check the 1st slide in each rack. I know this to be a common procedure with many histology professionals. The attitude can be left in your lab please. Thank you Cheryl Miller HT ASCP CM Histology Supervisor Physicians Laboratory Services Omaha, NE. 402 731 4148 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Monday, January 25, 2010 7:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Diff-Quik Thanks, John Kiernan, for your explanation of Romanovsky stains. Diff-Quik (please note the spelling) is the trademarked name of a staining sequence consisting of a fixative, eosin (Diff-Quik I), and an azure (Diff-Quik II), done in that order in three separate containers. I'm not sure who the trademark presently belongs to - it seems to change with the phases of the Moon. There are a number of generic equivalents, which in my personal experience all work as well as trademark Diff-Quik. For most ordinary pathology services, it isn't worthwhile to try to brew your own. I don't think I've seen bone marrow stained with such a sequence. Proper staining of bone marrows requires that the histotechnologist examine the slides under a microscope, a practice too many find abhorrent. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - All electronic data transmissions originating from or sent to Baptist Health Care Corporation (BHC) are subject to monitoring. This message along with
[Histonet] A blast from the past....
Dr Richmond, Thanks so much for that link to Vade Mecum. I've never before seen the book, but it has brought me back to my childhood, when I found a similar histotechnique book, Michael Guyer's Animal Micrology, in an antique shop my mom was browsing, some 44 years ago. This book was referenced all over that book, and in Gray's Microtomist's Formulary and Guide which I obtained later as my interest in histotechnique grew during high school, an interest kindled by that book and resulting in a rewarding and fascinating career. I feel like a kid again. Very cool. It's amazing how much mercury they slung around back then. Hell, I made my own Zenker's fluid in high school from those books, bet the waste went down the drain too. Jeff Silverman ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
http://www.nesle2005.republika.pl/T6vJjAE12x.html _ Hotmail: Trusted email with powerful SPAM protection. http://clk.atdmt.com/GBL/go/196390707/direct/01/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] honing compound
A colleague from a neighboring university is trying to find both fine and coarse honing compound for his knife sharpener. Does anyone have a good source? Thank you, Jennifer MacDonald ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
