[Histonet] Re: Decalcification and processing of large bone
I work with bone samples trimmed to approx 35mmx35mmx4mm. Our general procedure is: Fixation: untrimmed samples stored in 10% NBF @4°C until trimming. Slices placed in fresh 10% NBF for at least 12h. Decalcification Benchtop - 10% formic acid for 10 days (change soln every 3 days), rinse for minimum 1-2 hours. Then 10% EDTA solution for 2-3 weeks (change weekly). The time in formic acid is limited at 10 days to prevent excess damage to tissue/loss of nuclear staining. The EDTA is a chelating agent and is slower at decalcifying tissue but does little damage. Microwave processor - 3 days in 10% formic acid @35°C, rinse as above, then 3 days in 10% EDTA @35°C. For both methods I recommend placing a layer of empty cassettes at the bottom of you container to allow solution to circulate under samples. After the EDTA step, it is VITAL that samples are rinsed for at least 3 hours to remove any remainng salts. These precipitate during processing and make sectioning very difficult. You may need to leave samples longer in EDTA depending on their size. There are various methods in text books for determining decalcification end point. We have a MicroCT scanner so just do a quick xray scan to check our samples. Processing. We have a Sakura Tissue Tek 5. Samples processed for 5 hours each through 50%, 70%, 90% and 3x100% Ethanol/IMS. Then 4 hours x3 changes for chloroform, then 2 hours x4 changes of wax. Sectioning Make sure block are well chilled. Some samples may need soaking in water to get good sections. We trim in samples then place face down on a melting ice block whilst trimming in the other samples. This means our samples soak and chill at the same time. If you get small areas of mineral left, you can perform surface decal with 10% fromic acid. Hope this points you in the right direction. If you can, try and go to this years NSH convention. Lots of very helpful people there, especially the Hard Tissue Committee. Best of luck. Andrew Prior Histologist Smith Nephew Research Centre York Science Park UK andrew.pr...@smith-nephew.com -- Message: 21 Date: Wed, 24 Feb 2010 17:34:06 -0800 (PST) From: Victor Wong vhlw...@yahoo.com Subject: [Histonet] Decalcification and processing of large bone To: Histonet@lists.utsouthwestern.edu Message-ID: 347173.34928...@web52706.mail.re2.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Dear all, I am going to decal and process PIG femur and lumbar vertebrates. I only have experience on soft tissues. Anyone can suggest any protocols in decalcification and processing? Many thanks in advance. Cheers, Victor Confidentiality. This electronic transmission is strictly confidential to Smith Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. Smith Nephew UK Registered in England and Wales No.4421171 with registered office at 15 Adam Street, London WC2N 6LA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: [ Histonet ] Tris Buffer Preservative
I used to sterilze my TBS by autoclaving it. For aliquots I used TBS with 0,1% sodium acide. I never had any problems with bacteria; my 10x TBS concentrate was fine for weeks. - Original Message - From: amosbro...@gmail.com To: histonet@lists.utsouthwestern.edu Date: 24.02.2010 17:59:45 Subject: [Histonet] Tris Buffer Preservative Hi, I have been making up my own tris buffer for immunohistochemistry. It has been working swimmingly, with one exception. I have been doing some work on Salmonella, and apparently the researcher has been seeing bacteria in her immunofluorescent slides that she says are moving and seem to be viable. She wanted to culture all my reagents. So the primary and secondary antibody diluents (commercially purchased) came up clean, but the TBS ended up with a pretty healthy growing colony. Now since I don't do IHC overnight in an incubator, I don't think this is necessarily a catastrophy. (No one else has noticed this) It does seem to warrant further investigation though. So for the folks that make their own solutions up, what do you use as a preservative for your buffers and how much do you use. I haven't seen anything in any of the recipes I have found. I was thinking Sodium Azide, but it is really hazardous, and Wikipedia says it is actually explosive ( http://en.wikipedia.org/wiki/Sodium_azide). Biocare Medical's data sheet says they use less than 0.25% procyclin. (thanks for not hiding the ingredients Biocare, I love you for that!) Has anyone tried that? Any other suggestions would be welcome. Thanks, Amos ___ 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] Snap Freezing Tissue
Dear Laurie, We manufacture the Clini-RF Rapid Freezer =80 degs. C with options for a range of freezing methods. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abri...@brightinstruments.com Web Site: www.brightinstruments.com Skype: dazzle0 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: 24 February 2010 21:55 To: histonet@lists.utsouthwestern.edu Cc: Steve Pike Subject: [Histonet] Snap Freezing Tissue I was wondering how others are snap freezing tissue? We have been using a histobath (a refrigerator device that cools isopentane for freezing tissue), but it has died. I would love to have another Histobath, but they are no longer carried by the vendor that I originally bought mine from. Does anyone know where I can get either a new or refurbished Histobath (vendor calls are welcome)? I'm also interested in other freezing techniques. Laurie Colbert Huntington Hospital Pasadena, CA (626) 397-8620 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- BEGIN-ANTISPAM-VOTING-LINKS -- Teach SpamSniper if this mail (ID 755353) is spam: Spam:http://94.126.232.8/canit/b.php?i=755353m=c4db6e71fbd0c=s Not spam:http://94.126.232.8/canit/b.php?i=755353m=c4db6e71fbd0c=n Forget vote: http://94.126.232.8/canit/b.php?i=755353m=c4db6e71fbd0c=f -- END-ANTISPAM-VOTING-LINKS ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides
Hello Histonet, it has been a while (~10 months) since I posted a problem about uneven immuno-staining with specimen showing unstained circles after manual staining with HRP-polymere/DAB method; complete mail see below, response mails see Histonet archive (via website). Links to pictures I took: http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg The problem occured suddenly, without having changed any reagents or methods. Things I changed to avoid the unstained spots: *Adding 0,05% Tween 20 to TBS *Blocking peroxidase in coplin jar, not mounted in racks *Lowering antibody concentration which temporarily produced better results. After reviewing a big number of slides it showed up that most of the tissue affected was lung, liver and kidney which mostly means a lot of blood in the tissue when fixation in formalin starts. Erythrocytes, granulocytes and macrophages show a lot of endogenous and pseudoendogenous peroxidase activity. This is how it's done since then: Slides are taken from racks into a big coplin jar with 3% H2O2 diluted in distilled water (demineralized H2O). A slow magnetic stirrer on the bottom of the jar keeps the solution floating around the tissue, removing any O2 bubble that might appear. Slides then are remounted and rinsed a lot with TBS-T. Thank you for all your help, though it's a little late... V. Neubert, Germany - Original Message - From: histonet.nos...@vneubert.com To: histonet@lists.utsouthwestern.edu Date: 02.04.2009 18:12:18 Subject: [Histonet] Strange circles in IHC slides [...] http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg [...] So, has ever anyone experienced sth. like this? My conjugate control (every step except the antibody) was fine, nothing to be seen about DAB and no circles at all. I used Shandon single-use coverplates, sterile buffer, fresh antibody aliquots. Any idea? Thanks, V. Neubert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Snap Freezing Tissue
Hi Laurie, We distribute Bright's Clini-RF. We hear good reports about using 3MT NovecT Engineered Fluid HFE-7100 which is inert and has an excellent environmental, health and safety profile. Regards, Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com] Sent: Wednesday, February 24, 2010 4:55 PM To: histonet@lists.utsouthwestern.edu Cc: Steve Pike Subject: [Histonet] Snap Freezing Tissue I was wondering how others are snap freezing tissue? We have been using a histobath (a refrigerator device that cools isopentane for freezing tissue), but it has died. I would love to have another Histobath, but they are no longer carried by the vendor that I originally bought mine from. Does anyone know where I can get either a new or refurbished Histobath (vendor calls are welcome)? I'm also interested in other freezing techniques. Laurie Colbert Huntington Hospital Pasadena, CA (626) 397-8620 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Snap Freezing Tissue
I like this method too, and I would say it's one of the best. The only issue is high throughput. When freezing 100+ samples the floating tray becomes much more user friendly and believe it or not, the results are just as good. --- On Wed, 2/24/10, Adam . anonwu...@gmail.com wrote: From: Adam . anonwu...@gmail.com Subject: Re: [Histonet] Snap Freezing Tissue To: Andrea T. Hooper andreahoo...@rocketmail.com Cc: histonet@lists.utsouthwestern.edu, Laurie Colbert laurie.colb...@huntingtonhospital.com, Steve Pike ste...@mikronet.com Date: Wednesday, February 24, 2010, 10:31 PM I learned to freeze tissue by taking a beaker and filling it with 2-methylbutane and then placing that beaker in a liquid nitrogen bath. You then take place your cryomold, hold it using some long forceps, and place it on the top of the 2-methylbutane. It will freeze the entire thing in about 10 seconds. Once it's frozen, you dip the entire mold into the 2-methylbutane. Seems to work well for me. Adam ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Snap Freezing Tissue
Who supplies the 3MT Novec Engineered Fluid HFE-7100? Thanks Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael Williams Sent: Thursday, February 25, 2010 8:55 AM To: 'Laurie Colbert' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Snap Freezing Tissue Hi Laurie, We distribute Bright's Clini-RF. We hear good reports about using 3MT NovecT Engineered Fluid HFE-7100 which is inert and has an excellent environmental, health and safety profile. Regards, Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com] Sent: Wednesday, February 24, 2010 4:55 PM To: histonet@lists.utsouthwestern.edu Cc: Steve Pike Subject: [Histonet] Snap Freezing Tissue I was wondering how others are snap freezing tissue? We have been using a histobath (a refrigerator device that cools isopentane for freezing tissue), but it has died. I would love to have another Histobath, but they are no longer carried by the vendor that I originally bought mine from. Does anyone know where I can get either a new or refurbished Histobath (vendor calls are welcome)? I'm also interested in other freezing techniques. Laurie Colbert Huntington Hospital Pasadena, CA (626) 397-8620 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Snap Freezing Tissue
It can be purchased directly from 3M. You may also find some local suppliers. Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Houston, Ronald [mailto:ronald.hous...@nationwidechildrens.org] Sent: Thursday, February 25, 2010 9:42 AM To: Michael Williams; Laurie Colbert Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Snap Freezing Tissue Who supplies the 3MT Novec Engineered Fluid HFE-7100? Thanks Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael Williams Sent: Thursday, February 25, 2010 8:55 AM To: 'Laurie Colbert' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Snap Freezing Tissue Hi Laurie, We distribute Bright's Clini-RF. We hear good reports about using 3MT NovecT Engineered Fluid HFE-7100 which is inert and has an excellent environmental, health and safety profile. Regards, Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com] Sent: Wednesday, February 24, 2010 4:55 PM To: histonet@lists.utsouthwestern.edu Cc: Steve Pike Subject: [Histonet] Snap Freezing Tissue I was wondering how others are snap freezing tissue? We have been using a histobath (a refrigerator device that cools isopentane for freezing tissue), but it has died. I would love to have another Histobath, but they are no longer carried by the vendor that I originally bought mine from. Does anyone know where I can get either a new or refurbished Histobath (vendor calls are welcome)? I'm also interested in other freezing techniques. Laurie Colbert Huntington Hospital Pasadena, CA (626) 397-8620 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides
To me it seems that the sections after being picked from the water bath were not completely drained and the dewaxing process was incomplete in a way that the round areas kept certain amount of paraffin wax that prevented the reagents reactions. The fact that the areas are round are an indication that water was involved, since water always leave a round imprint, due to its surface tension. I would suggest that you dewax the sections with a 2% aq. solution of dish washer soap. Dewaxing with xylene sections containing water will be incomplete because it does not mix completely with water but the detergent will mix with the water and will better remove the paraffin.René J. --- On Thu, 2/25/10, histonet.nos...@vneubert.com histonet.nos...@vneubert.com wrote: From: histonet.nos...@vneubert.com histonet.nos...@vneubert.com Subject:[Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides To: histonet@lists.utsouthwestern.edu Date: Thursday, February 25, 2010, 8:34 AM Hello Histonet, it has been a while (~10 months) since I posted a problem about uneven immuno-staining with specimen showing unstained circles after manual staining with HRP-polymere/DAB method; complete mail see below, response mails see Histonet archive (via website). Links to pictures I took: http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg The problem occured suddenly, without having changed any reagents or methods. Things I changed to avoid the unstained spots: *Adding 0,05% Tween 20 to TBS *Blocking peroxidase in coplin jar, not mounted in racks *Lowering antibody concentration which temporarily produced better results. After reviewing a big number of slides it showed up that most of the tissue affected was lung, liver and kidney which mostly means a lot of blood in the tissue when fixation in formalin starts. Erythrocytes, granulocytes and macrophages show a lot of endogenous and pseudoendogenous peroxidase activity. This is how it's done since then: Slides are taken from racks into a big coplin jar with 3% H2O2 diluted in distilled water (demineralized H2O). A slow magnetic stirrer on the bottom of the jar keeps the solution floating around the tissue, removing any O2 bubble that might appear. Slides then are remounted and rinsed a lot with TBS-T. Thank you for all your help, though it's a little late... V. Neubert, Germany - Original Message - From: histonet.nos...@vneubert.com To: histonet@lists.utsouthwestern.edu Date: 02.04.2009 18:12:18 Subject: [Histonet] Strange circles in IHC slides [...] http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg [...] So, has ever anyone experienced sth. like this? My conjugate control (every step except the antibody) was fine, nothing to be seen about DAB and no circles at all. I used Shandon single-use coverplates, sterile buffer, fresh antibody aliquots. Any idea? Thanks, V. Neubert ___ 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] Histology position at Indianapolis VA Medical center
Hi, There is a great hospital based Histotechnologists position now available in the great midwestern city of Indianapolis, Indiana. Follow the link provided: http://jobsearch.usajobs.gov/search.aspx?q=KB-10-SRC-320328where=brd=3876vw=bFedEmp=NFedPub=Yx=55y=14 Sincerely, Nick Frain, AP supervisor Roudebush VA Medical Center Indianapolis, IN 46202 O-317-988-2217 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Snap Freezing Tissue
In response to the interested in other freezing techniques part of Laurie's query: We snap freeze by throwing a cyrovial of tissue (or a tissue piece wrapped in foil) into some liquid nitrogen for a couple minutes (vials will float, foil-wrapped pieces will sink). For controlled freezing of OCT-embedded tissues, I've frozen isopentane by pouring some in a liquid-nitrogen proof shallow container (such as a pipet-tip box lid) and floating it on the liquid nitrogen til frozen. I guess this works if you're processing a relatively small number of samples at a time, which, if you work in a hospital, sounds like you may not be. Additionally, we've made alcohol-dry ice slurries in styrofoam containers in which to freeze OCT-embedded tissues. Regards, Merced --On Thursday, February 25, 2010 10:12 AM -0500 Michael W illiams mjwilli...@scilogex.com wrote: It can be purchased directly from 3M. You may also find some local suppliers. Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Houston, Ronald [mailto:ronald.hous...@nationwidechildrens.org] Sent: Thursday, February 25, 2010 9:42 AM To: Michael Williams; Laurie Colbert Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Snap Freezing Tissue Who supplies the 3MT Novec Engineered Fluid HFE-7100? Thanks Ronnie Houston Anatomic Pathology Manager Nationwide Children's Hospital Columbus OH 43205 (614) 722 5450 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael Williams Sent: Thursday, February 25, 2010 8:55 AM To: 'Laurie Colbert' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Snap Freezing Tissue Hi Laurie, We distribute Bright's Clini-RF. We hear good reports about using 3MT NovecT Engineered Fluid HFE-7100 which is inert and has an excellent environmental, health and safety profile. Regards, Michael Williams mjwilli...@scilogex.com SCILOGEX, LLC TF: (877) SCILOGEX T: (860) 828-5614 F: (860) 828-5389 www.SCILOGEX.com -Original Message- From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com] Sent: Wednesday, February 24, 2010 4:55 PM To: histonet@lists.utsouthwestern.edu Cc: Steve Pike Subject: [Histonet] Snap Freezing Tissue I was wondering how others are snap freezing tissue? We have been using a histobath (a refrigerator device that cools isopentane for freezing tissue), but it has died. I would love to have another Histobath, but they are no longer carried by the vendor that I originally bought mine from. Does anyone know where I can get either a new or refurbished Histobath (vendor calls are welcome)? I'm also interested in other freezing techniques. Laurie Colbert Huntington Hospital Pasadena, CA (626) 397-8620 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet - Confidentiality Notice: The following mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. ___ 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
[Histonet] NBS/NIST Thermometers
Does anyone know how frequently you have to certify your NIST Thermometers (the thermometer you use to calibrate all of the thermometers in the laboratory), including a reference. The certifiicate for my NIST Thermometer does not document the next time it needs to be calibrated. Thank you, Ann Angelo ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Control slides for Flag-Tag IHC
Folks, I am looking for a control for some Flag-tag IHC I am doing on FFPE tissue. I was wondering if someone could share some slides off of a block of flag transfected cells. THANKS!! Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Director Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. DE-360 (Please note new location) Seattle WA 98109-1024 206-667-6119 jhabe...@fhcrc.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Control tissue for Fluoro Jade
Hello Histonet, I am looking for a positive control for the Fluoro Jade Stain consisting of rodent brain tissue demonstrating neuronal necrosis in the hippocampus caused by administration of trimethyltin. Does anyone have access to a stock of controls or can anyone tell me who to contact. I have contacted the AFIP and NSH but neither were able to help. Any information would be greatly appreciated. Thank-you, Jonilyn Van Fleet, HT (ASCP) The Dow Chemical Company 989-636-3539 jvanfl...@dow.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] NBS/NIST Thermometers
I am in NY State and they just told me yesterday that we need to do it annually. Loralee McMahon, HTL (ASCP) Immunohistochemistry Supervisor Strong Memorial Hospital Department of Surgical Pathology (585) 275-7210 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of thisis...@aol.com [thisis...@aol.com] Sent: Thursday, February 25, 2010 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NBS/NIST Thermometers Does anyone know how frequently you have to certify your NIST Thermometers (the thermometer you use to calibrate all of the thermometers in the laboratory), including a reference. The certifiicate for my NIST Thermometer does not document the next time it needs to be calibrated. Thank you, Ann Angelo ___ 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] Re: Snap Freezing Tissue
In the last two or three years I and others have posted a number of times on Histonet about the Histobath (which is no longer in manufacture), Bright's Clini-RF, and 3M's NovecT Engineered Fluid HFE-7100. You can access these posts in the Archives, through Google if you wish. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] haematoxylin
Hello everyone! Does anyone know where I can get more information about haematoxylin-haematein-oxyhaematein chemistry? I have a problem with my Harris haematoxylin modification (sodium iodate as oxidant). When I use a smaller amount of sodium iodate (about 0,1g per 1g of haematoxylin) I have a huge problem with precipitation. I don't know if this is because of the unoxidized haematoxylin in staining solution or because of something else. Any help or suggestions are welcomed! Thanks in advance! Peter ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] voice recognition
Hello, Can anyone using voice recognition with Cerner Millennium tell me what system they are using and how they like it? Thanks, Jan Mahoney Omaha, NE Sponsored by Catholic Health Initiatives and Immanuel Health Systems, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] haematoxylin
0.1G should give you an approximately half-oxidized hamatoxylin solution along the lines of Baker's haematal-8 or haematal-16, or Gill's haemalum. It should keep well. Precipitation must be due to something else you're doing. For the chemistry a good starting point is Conn's Biological Stains (10th ed, 2002). for information, see http://biologicalstaincommission.org and click on Publications. The current issue of Biotechnic Histochemistry (Feb. 2010) is a special issue devoted to haematoxylin (Vol 85 No. 1). Last year the same journal (Vol 84 No. 4, pp. 159-177) carried an 18-page paper, Nuclear staining with alum hematoxylin by Bryan Llewellyn, which includes recipes for scores of haemalum mixtures and has much other interesting information. You can review the contents of the journal at http://informahealthcare.com/loi/bih. If your institution's library subscribes to the Informa Healthcare package of journals, you'll be able to download PDF files of articles - they go right back to 1925. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: Hana Peter hana...@gmail.com Date: Thursday, February 25, 2010 17:16 Subject: [Histonet] haematoxylin To: histonet@lists.utsouthwestern.edu Hello everyone! Does anyone know where I can get more information about haematoxylin-haematein-oxyhaematein chemistry? I have a problem with my Harris haematoxylin modification (sodium iodate as oxidant). When I use a smaller amount of sodium iodate (about 0,1g per 1g of haematoxylin) I have a huge problem with precipitation. I don't know if this is because of the unoxidized haematoxylin in staining solution or because of something else. Any help or suggestions are welcomed! Thanks in advance! Peter ___ 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
