....IMEB, INC. had a clear paraffin at the NSH meeting in birmingham. They are at www.IMEBINC.com E-van
________________________________________ From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu [histonet-requ...@lists.utsouthwestern.edu] Sent: Wednesday, October 14, 2009 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 71, Issue 14 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. image pro plus (Sarka Lhotak) 2. price for plastic embedding (Nancy W. Troiano) 3. Clear Paraffin? (Monfils, Paul) 4. Transporting Formalin (Feher, Stephen) 5. New Lead Tech/Assistant Manager position available in NJ (Brian- Prometheus) 6. how to fix EGFP for proteomics (Lilja Thoenes) 7. Pathos Tissue Processor vs Tissue Xpress (Kiranjit Grewal) 8. hematoxylin and nova red staining (del phillips) 9. looking for calibration tools for Optimax plus stainer (Leroy Brown) 10. Re: looking for calibration tools for Optimax plus stainer (Patti Loykasek) 11. Isotype background (Adam .) 12. RE: Automated H&E stainers (Cathy) 13. RE: Transporting Formalin (DELIA GARCIA) 14. lid gasket for processor (Diana McCaig) 15. pkh (Melissa Mazan) 16. RE: lid gasket for processor (Rathborne, Toni) 17. Fw: Pathos Tissue Processor vs Tissue Xpress (Rene J Buesa) 18. Re: Isotype background (Merced M Leiker) 19. RE: Fw: Pathos Tissue Processor vs Tissue Xpress (Rene J Buesa) 20. HerCep (kk...@bidmc.harvard.edu) 21. unsubscribe, please (susan bryant) 22. RELIA Histology Job Alert Histotechnologist needed in Atlanta (Pam Barker) 23. Re: unsubscribe, please (Emily Sours) 24. Re: unsubscribe, please (Peter Carroll) ---------------------------------------------------------------------- Message: 1 Date: Tue, 13 Oct 2009 13:30:14 -0400 From: "Sarka Lhotak" <lhot...@mcmaster.ca> Subject: [Histonet] image pro plus To: histonet@lists.utsouthwestern.edu Message-ID: <web-273091...@cgpsrv2.cis.mcmaster.ca> Content-Type: text/plain;charset=utf-8 Hi Hatem, to separate RGB colours, go to Process/ Colour channel/ Extract. This will give you each channel in B&W, i.e. each pixel with intensity from 0 to 255 for Red, Blue and Green component of your image. There must be many ways how to do intensity. One that would be possible is: go to Process/ Pseudocolour/ Select number of divisions (bins). Intensities will then be divided into bins, say 0 to 10, 11 to 20, 21 to 30 etc. all the way to 255. And then look in areas, it will give you number of pixels for each range of intensities you selected in Divisions. Hope it helps, Sarka Lhotak, PhD McMaster University, Hamilton ------------------------------ Message: 2 Date: Tue, 13 Oct 2009 15:26:34 -0400 From: "Nancy W. Troiano" <nancy.troi...@yale.edu> Subject: [Histonet] price for plastic embedding To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20091013152448.0145d...@email.med.yale.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Our lab prices for plastic embedding are dependent on many variables, e.g. size of bone tissue, implant/nonimplant, services required (such as embed only, embed, cut, etc.). You can contact me for pricing for work done for outside institutions. ------------------------------ Message: 3 Date: Tue, 13 Oct 2009 15:58:46 -0400 From: "Monfils, Paul" <pmonf...@lifespan.org> Subject: [Histonet] Clear Paraffin? To: <histonet@lists.utsouthwestern.edu> Message-ID: <4ebff65383b74d49995298c4976d1d5e03835...@lsriexch1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I work in a core histology facility where I receive all kinds of specimens and embedded blocks from many different sources. Occasionally I receive blocks that are embedded in a clear (transparent, not white) embedding wax. Does anyone know what this product is? ------------------------------ Message: 4 Date: Tue, 13 Oct 2009 16:23:10 -0400 From: "Feher, Stephen" <sfe...@cmc-nh.org> Subject: [Histonet] Transporting Formalin To: <histonet@lists.utsouthwestern.edu> Message-ID: <73a7ed895ee0c24d9267ed814911df190fdb2...@exchange.cmc-nh.org> Content-Type: text/plain; charset="us-ascii" Hi gang, A question came up in a committee meeting today surrounding how other institutions are transporting quantities of 10% NBF from a main facility to outlying hospitals and outpatient clinics. The quantities in question ranged from 5 - 20 gallons of formalin in sealed carboy type containers. Does a main hospital or lab distribute the formalin to outlying facilities or are outlying facilities having to order direct from a supplier and bill the main facility? Some places just order it and transport via courier. This may be restricted due to various State EPA rules. How are you all doing this? Thanks, Steve Stephen A. Feher, MS, SCT (ASCP) Pathology Supervisor Catholic Medical Center 100 McGregor Street Manchester, NH 03102 603-663-6707 sfe...@cmc-nh.org <mailto:sfe...@cmc-nh.org> ------------------------------ Message: 5 Date: Tue, 13 Oct 2009 16:52:32 -0400 From: "Brian- Prometheus" <br...@prometheushealthcare.com> Subject: [Histonet] New Lead Tech/Assistant Manager position available in NJ To: <histonet@lists.utsouthwestern.edu> Message-ID: <001901ca4c47$15e77600$41b662...@com> Content-Type: text/plain; charset="us-ascii" New Opening with a lab in Paramus Day shift 5am to 1:30pm or 6:00a to 2:30pm Stresses quality over quantity Prefers ASCP certification and NY state licensure Someone who can multitask, detail oriented IHC experience a plus Salary will commensurate upon experience Minimum 10 yrs exp Lab is growing, consistently acquiring new clients. Been in business about a year Please let me know if you may be interested! Brian Feldman Principal Prometheus Healthcare Office 301-693-9057 Fax 301-368-2478 br...@prometheushealthcare.com <mailto:br...@prometheushealthcare.com> www.prometheushealthcare.com <http://www.prometheushealthcare.com/> *** Stay up to date on the newest positions and healthcare trends nationwide on Twitter!*** http://twitter.com/PrometheusBlog ------------------------------ Message: 6 Date: Fri, 9 Oct 2009 16:01:23 +0100 From: Lilja Thoenes <liljathoe...@rcsi.ie> Subject: [Histonet] how to fix EGFP for proteomics To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <54e3312540de1547a410fa8a333ac8f00223fa4...@rcsiexchange.rcsi-internal.ie> Content-Type: text/plain; charset="us-ascii" Hi , I plan isolate GFPtagged cells from cryosections via laser microdissection for proteomics but still I am struggling with not loosing the GFP signal. Does anyone has a good idea how to fix the signal so that afterwards proteomics is still possible? Thanks a lot, Lilja Lilja Thoenes Dept. of Neurodegeneration and Physiology Royal College of Surgeons in Ireland (RCSI) 123 St. Stephens Green, Dublin 2, Ireland Office: +353 1 402 2794 Fax: +353 1 402 2447 ------------------------------ Message: 7 Date: Sun, 11 Oct 2009 21:07:39 -0700 From: "Kiranjit Grewal" <kira...@sbcglobal.net> Subject: [Histonet] Pathos Tissue Processor vs Tissue Xpress To: <histonet@lists.utsouthwestern.edu> Message-ID: <277641.14522...@smtp101.sbc.mail.sp1.yahoo.com> Content-Type: text/plain; charset="us-ascii" Hi, Anybody has anything to share regarding Pathos both good and bad. We are planning to get one. Also please share if you have any experience with Tissue Xpress. Thanks, Histotech in CA ------------------------------ Message: 8 Date: Mon, 12 Oct 2009 10:36:08 -0500 From: "del phillips" <dphill...@vetmed.lsu.edu> Subject: [Histonet] hematoxylin and nova red staining To: <histonet@lists.utsouthwestern.edu> Cc: dphillip...@juno.com Message-ID: <000001ca4b52$2f60e080$8e22a1...@lsu.edu> Content-Type: text/plain; charset="us-ascii" If hematoxylin is place on a slide before the nova red is it too late to add the nova red after the hematoxylin? Thanks ------------------------------ Message: 9 Date: Tue, 13 Oct 2009 14:10:39 -0700 From: Leroy Brown <rhbro...@histocs.com> Subject: [Histonet] looking for calibration tools for Optimax plus stainer To: <histonet@lists.utsouthwestern.edu> Message-ID: <mailman.0.1255539601.17299.histo...@lists.utsouthwestern.edu> Content-Type: text/plain; charset="iso-8859-1"; Does anyone have a set of calibration tools needed to calibrate my BioGenex Optimax stainer. Seems they no longer make these or sell time?? thanks LeRoy Brown Ht(ASCP) HTL www.histocs.com 360-966-7300 ------------------------------ Message: 10 Date: Tue, 13 Oct 2009 14:46:37 -0700 From: Patti Loykasek <ployka...@phenopath.com> Subject: Re: [Histonet] looking for calibration tools for Optimax plus stainer To: <histonet@lists.utsouthwestern.edu> Message-ID: <c6fa434d.1b22f%ployka...@phenopath.com> Content-Type: text/plain; charset="US-ASCII" In reference to LeRoy's last statement below- if anyone is selling time please put me on the list for the maximum allowable! I haven't been able to figure out how to cram more hours in a day! And it's only Tuesday. Patti Loykasek > Does anyone have a set of calibration tools needed to calibrate my BioGenex > Optimax stainer. Seems they no longer make these or sell time?? > > thanks > > LeRoy Brown Ht(ASCP) HTL > > www.histocs.com > > 360-966-7300 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. ------------------------------ Message: 11 Date: Tue, 13 Oct 2009 16:48:46 -0500 From: "Adam ." <anonwu...@gmail.com> Subject: [Histonet] Isotype background To: histonet@lists.utsouthwestern.edu Message-ID: <858249120910131448v26622f8dtd5599dbbf64e6...@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 Hi all, I am trying some IHC, and I am having a peculiar problem. Like I expect, my antibody of interest (anti-mouse goat polyclonal) stains nonspecifically at high concentrations (10 ug / ml) but as I titer it down (3 ug / mL), it seems to stain relatively specifically the cells I think it should stain. However, at 3 ug / mL, my isotype goat IgG stains nearly everything. Here is my protocol 1) Block in 3% H2O2 for 10'. Wash. 2) Block in 10% donkey serum for 1 hr. Wash. 3) Block in avidin 15', wash, biotin 15', wash (Vector Labs blocking kits) 4) Incubate with primary / isotype overnight at 4C in 2% donkey serum. Wash. 5) Incubate with secondary (biotinylated donkey anti-goat, cross adsorbed to mouse) in 2% donkey serum for 1 hr at room temp. Wash. 6) Incubate with strepavidin HRP in TBS-T for 30'. Wash. 7) Incubate with DAB+ (Dako) for 5'. For isotype, I am using Chrompure Goat IgG, whole molecule from Jackson. Some people have suggested that I just do away with isotypes altogether and use a no primary control instead. I think there is some merit to this idea, but I still think my issue might be indicative of a larger technical problem in my staining protocol. Thanks, Adam ------------------------------ Message: 12 Date: Tue, 13 Oct 2009 19:10:08 -0700 From: "Cathy" <cf...@telus.net> Subject: RE: [Histonet] Automated H&E stainers To: "'Bauer, Karen L.'" <bauer.ka...@mayo.edu>, "'Connolly, Brett M'" <brett_conno...@merck.com>, <histonet@lists.utsouthwestern.edu> Message-ID: <9efa5fe9872a49718a169c823ad49...@your8ba846406f> Content-Type: text/plain; charset="us-ascii" Have you had any problems with the Coverslipper? We purchased the Prisma with linked Coverslipper last March and have had on going problems with the Coverslipper jamming with error code -Motor ejector error E15. We have contacted Somagen and were told that this is normal. It is happening at least once a day and sometimes more. We love the stainer and if the Coverslipper wasn't jamming we would love it just as much. What do you do with the coverslipped slides? We find that they don't dry as quickly as they did on the older film Coverslipper. We don't have a vented sign out area so they are staying on the carousel until they are mostly dry. Cathy -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bauer, Karen L. Sent: Monday, October 12, 2009 5:42 AM To: Connolly, Brett M; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Automated H&E stainers Brett, We purchased the Tissue Tek PRISMA with the attached film coverslipper a year ago and LOVE IT!! Was a little nervous switching from the glass coverslips to the film, but the docs love the slides and we have had no problems. Very easy to use and we like the fact that we can file the slides the same day without having to wait for them to "dry". Karen Karen L. Bauer HTL (ASCP) Histology Section Chief Department of Pathology - Luther Hospital Luther Midelfort - Mayo Health System 715-838-3205 bauer.ka...@mayo.edu -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M Sent: Friday, October 09, 2009 8:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated H&E stainers Which one do you like?...pro&cons? Thanks, Brett Brett M. Connolly, Ph.D. Research Fellow, Imaging Dept. Merck & Co., Inc. PO Box 4, WP-44K West Point, PA 19486 tel. 215-652-2501 fax. 215-993-6803 brett_conno...@merck.com Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu - direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ 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 ------------------------------ Message: 13 Date: Tue, 13 Oct 2009 21:52:57 CDT From: "DELIA GARCIA" <deliad...@yahoo.com> Subject: RE: [Histonet] Transporting Formalin To: "Feher, Stephen" <sfe...@cmc-nh.org>,histonet@lists.utsouthwestern.edu Message-ID: <mailman.1.1255539601.17299.histo...@lists.utsouthwestern.edu> Content-Type: text/plain; charset="iso-8859-1" We have various shipping addresses with our supplier. Which allows us to order directly for our clients. We incur all costs including the associated Hazardous Materials handling fee. We also use courier for in-town clients. Hope this helps. Delia Garcia Lead Histotechnologist Tucson, AZ -----Original Message----- Date: Tuesday, October 13, 2009 1:36:33 pm To: <histonet@lists.utsouthwestern.edu> From: "Feher, Stephen" <sfe...@cmc-nh.org> Subject: [Histonet] Transporting Formalin Hi gang, A question came up in a committee meeting today surrounding how other institutions are transporting quantities of 10% NBF from a main facility to outlying hospitals and outpatient clinics. The quantities in question ranged from 5 - 20 gallons of formalin in sealed carboy type containers. Does a main hospital or lab distribute the formalin to outlying facilities or are outlying facilities having to order direct from a supplier and bill the main facility? Some places just order it and transport via courier. This may be restricted due to various State EPA rules. How are you all doing this? Thanks, Steve Stephen A. Feher, MS, SCT (ASCP) Pathology Supervisor Catholic Medical Center 100 McGregor Street Manchester, NH 03102 603-663-6707 sfe...@cmc-nh.org <mailto:sfe...@cmc-nh.org> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 14 Oct 2009 06:33:32 -0400 From: "Diana McCaig" <dmcc...@ckha.on.ca> Subject: [Histonet] lid gasket for processor To: <histonet@lists.utsouthwestern.edu> Message-ID: <dcfd9e6a390e294aaf3a2561cd32e5c40d577...@ckhamail1.ckha.on.ca> Content-Type: text/plain; charset="us-ascii" Our lid gasket seemed to have grown overnight and expanded considerable in length. It is difficult to re-seat it back in. It appears too loose and when the processor cycles through a station and we open the lid, it is laying on the edge of the retort. Does anyone have any ideas what we can do until a new gasket can be ordered? Diana ------------------------------ Message: 15 Date: Wed, 14 Oct 2009 08:22:21 -0400 From: Melissa Mazan <melissa.ma...@tufts.edu> Subject: [Histonet] pkh To: histonet@lists.utsouthwestern.edu Message-ID: <4ad5c27d.5010...@tufts.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hi all, We have collected a specific lung cell via FACS from mice, stained the cells with PKH, and then returned the cells to congenic mice via 1. tracheal injection or 2. tail vein injection. We can see the PKH positive cells in the recipient lungs on microscopy, but we are having a real problem costaining these cells with anything else such as vimentin - has anyone else tried this approach? Melissa -- Melissa R. Mazan, DVM, Diplomate ACVIM Associate Professor and Director of Equine Sports Medicine Department of Clinical Sciences Tufts Cummings School of Veterinary Medicine 200 Westborough Road North Grafton,MA 01536 tel: 508-839-5395 fax: 508-839-7903 email: melissa.ma...@tufts.edu ------------------------------ Message: 16 Date: Wed, 14 Oct 2009 09:26:52 -0400 From: "Rathborne, Toni" <trathbo...@somerset-healthcare.com> Subject: RE: [Histonet] lid gasket for processor To: "Diana McCaig" <dmcc...@ckha.on.ca>, <histonet@lists.utsouthwestern.edu> Message-ID: <e78340c766a5284d999f5f5891ddf8900baf8...@smcmail.somerset-healthcare.com> Content-Type: text/plain; charset="utf-8" You might be able to shrink it a little by putting the gasket in the refrigerator for a while. -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Diana McCaig Sent: Wednesday, October 14, 2009 6:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lid gasket for processor Our lid gasket seemed to have grown overnight and expanded considerable in length. It is difficult to re-seat it back in. It appears too loose and when the processor cycles through a station and we open the lid, it is laying on the edge of the retort. Does anyone have any ideas what we can do until a new gasket can be ordered? Diana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. -------------------------------------------------------------- Somerset Medical Center is the recipient of the 2009 Orthopedic Surgery Excellence Award(tm) from HealthGrades, the nation's leading health care ratings company. Visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for news, event listings, health information and more. Join the Discussion: Facebook: www.somersetmedicalcenter.com/fb Twitter: www.twitter.com/SomersetMedCtr ------------------------------ Message: 17 Date: Wed, 14 Oct 2009 06:30:14 -0700 (PDT) From: Rene J Buesa <rjbu...@yahoo.com> Subject: [Histonet] Fw: Pathos Tissue Processor vs Tissue Xpress To: histonet@lists.utsouthwestern.edu Message-ID: <605622.63517...@web65713.mail.ac4.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 --- On Wed, 10/14/09, Rene J Buesa <rjbu...@yahoo.com> wrote: From: Rene J Buesa <rjbu...@yahoo.com> Subject: Pathos Tissue Processor vs Tissue Xpress To: "Kiranjit Grewal" <kira...@sbcglobal.net> Date: Wednesday, October 14, 2009, 9:28 AM Under separate cover I am sending you an article where both instruments (as well as some others) are compared in?reurn/investment and effectiveness. Ren? J. --- On Mon, 10/12/09, Kiranjit Grewal <kira...@sbcglobal.net> wrote: From: Kiranjit Grewal <kira...@sbcglobal.net> Subject: [Histonet] Pathos Tissue Processor vs Tissue Xpress To: histonet@lists.utsouthwestern.edu Date: Monday, October 12, 2009, 12:07 AM Hi, Anybody has anything to share regarding Pathos both good and bad. We are planning to get one. Also please share if you have any experience with Tissue Xpress. Thanks, Histotech in CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 14 Oct 2009 10:20:41 -0400 From: Merced M Leiker <lei...@buffalo.edu> Subject: Re: [Histonet] Isotype background To: "Adam ." <anonwu...@gmail.com>, histonet@lists.utsouthwestern.edu Message-ID: <31878536799288738d402...@cdywxp1931.ad.med.buffalo.edu> Content-Type: text/plain; charset=us-ascii; format=flowed I wouldn't get rid of the isotype control altogether; it's fulfilling it's purpose in trying to tell you something. Have you tried a no-primary control in parallel with your isotype control to see what the secondary is binding to - the tissue or bound goat IgG? What type of tissue is it? Maybe the goat IgG is binding to Fc receptors in the tissue. Try an Fc block. Or try a different goat IgG or one from a different company. But it may not even be a big issue; your goat IgG is not giving you the specific stain pattern that you are observing with your primary, right? It's just giving a lot of background? Instead of spending a lot of time troubleshooting the high background issue with your isotype control, it's already succeeded in telling you that your primary is specific, which is what you ultimately wanted to know, so you could just take that for what it's worth and go from there... Just my two cents' worth! Regards, Merced --On Tuesday, October 13, 2009 4:48 PM -0500 "Adam ." <anonwu...@gmail.com> wrote: > Hi all, > > I am trying some IHC, and I am having a peculiar problem. Like I expect, > my antibody of interest (anti-mouse goat polyclonal) stains > nonspecifically at high concentrations (10 ug / ml) but as I titer it > down (3 ug / mL), it seems to stain relatively specifically the cells I > think it should stain. However, at 3 ug / mL, my isotype goat IgG stains > nearly everything. > > Here is my protocol > 1) Block in 3% H2O2 for 10'. Wash. > 2) Block in 10% donkey serum for 1 hr. Wash. > 3) Block in avidin 15', wash, biotin 15', wash (Vector Labs blocking kits) > 4) Incubate with primary / isotype overnight at 4C in 2% donkey serum. > Wash. 5) Incubate with secondary (biotinylated donkey anti-goat, cross > adsorbed to mouse) in 2% donkey serum for 1 hr at room temp. Wash. > 6) Incubate with strepavidin HRP in TBS-T for 30'. Wash. > 7) Incubate with DAB+ (Dako) for 5'. > > For isotype, I am using Chrompure Goat IgG, whole molecule from Jackson. > Some people have suggested that I just do away with isotypes altogether > and use a no primary control instead. I think there is some merit to this > idea, but I still think my issue might be indicative of a larger > technical problem in my staining protocol. > > Thanks, > Adam > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 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. ------------------------------ Message: 19 Date: Wed, 14 Oct 2009 07:22:38 -0700 (PDT) From: Rene J Buesa <rjbu...@yahoo.com> Subject: RE: [Histonet] Fw: Pathos Tissue Processor vs Tissue Xpress To: Donna Willis <do...@milestonemed.com> Cc: histonet@lists.utsouthwestern.edu Message-ID: <158309.9167...@web65701.mail.ac4.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Donna: Thank you for your comment. As you write if the Pathos Classic is still available what I wrote is still valid. Unfortunately when somebody writes an article the conclusions remain valid as long as the subjects don't change, but the approach remains. As you can see I have forwarded your comments and my answer for all to read in HistoNet so they can benefit of it. As to going to Kalamazzo I really appreciate your invitation but I seldom travel ("retired with a fixed income", you the drill!). Regards Ren? J. --- On Wed, 10/14/09, Donna Willis <do...@milestonemed.com> wrote: From: Donna Willis <do...@milestonemed.com> Subject: RE: [Histonet] Fw: Pathos Tissue Processor vs Tissue Xpress To: "Rene J Buesa" <rjbu...@yahoo.com> Date: Wednesday, October 14, 2009, 9:58 AM Rene, There are now 2 Pathos units available to customers.? The Pathos Classic and the Pathos Delta.? You article was written before the Delta was on the market.? Please understand that giving this article to customers may not give them a true picture of the unit they are looking to purchase.? If you are interested in coming to the Milestone Kalamazoo office to experience the whole Milestone instrument line, we would love to have you. Sincerely, Donna Willis, HT/HTL(ASCP) North American Application Manager Milestone Medical (866) 995-5300 toll free (269) 488-4040 fax www.milestonemed.com Helping Patients -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, October 14, 2009 8:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fw: Pathos Tissue Processor vs Tissue Xpress --- On Wed, 10/14/09, Rene J Buesa <rjbu...@yahoo.com> wrote: From: Rene J Buesa <rjbu...@yahoo.com> Subject: Pathos Tissue Processor vs Tissue Xpress To: "Kiranjit Grewal" <kira...@sbcglobal.net> Date: Wednesday, October 14, 2009, 9:28 AM Under separate cover I am sending you an article where both instruments (as well as some others) are compared in?reurn/investment and effectiveness. Ren? J. --- On Mon, 10/12/09, Kiranjit Grewal <kira...@sbcglobal.net> wrote: From: Kiranjit Grewal <kira...@sbcglobal.net> Subject: [Histonet] Pathos Tissue Processor vs Tissue Xpress To: histonet@lists.utsouthwestern.edu Date: Monday, October 12, 2009, 12:07 AM Hi, Anybody has anything to share regarding Pathos both good and bad. We are planning to get one. Also please share if you have any experience with Tissue Xpress. Thanks, Histotech in CA _______________________________________________ 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 ------------------------------ Message: 20 Date: Wed, 14 Oct 2009 10:25:51 -0400 From: kk...@bidmc.harvard.edu Subject: [Histonet] HerCep To: histonet@lists.utsouthwestern.edu Message-ID: <1453b63a4be95441aa65f577cee7b6e104e1699...@evs5ccr.its.caregroup.org> Content-Type: text/plain; charset=us-ascii Does anyone have any experience using the Dako HerCep kit? And do you have any problems with cytoplasmic staining? ------------------------------ Message: 21 Date: Wed, 14 Oct 2009 11:24:44 -0400 From: susan bryant <sbry...@labpath.com> Subject: [Histonet] unsubscribe, please To: histonet@lists.utsouthwestern.edu Message-ID: <4ad5ed3c.20...@labpath.com> Content-Type: text/plain; charset=ISO-8859-1; format=flowed I wish to unsubscribe and return in 6 weeks. Thank you, Susan E. Bryant Knoxville Dermatopathology ------------------------------ Message: 22 Date: Wed, 14 Oct 2009 11:42:07 -0400 From: "Pam Barker" <rel...@earthlink.net> Subject: [Histonet] RELIA Histology Job Alert Histotechnologist needed in Atlanta To: "'Histonet'" <histonet@lists.utsouthwestern.edu> Message-ID: <e1my5zm-0005zb...@elasmtp-curtail.atl.sa.earthlink.net> Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters! I have a new position that I want to let everyone know about. I am assisting a client located in Atlanta GA that is in need of a histotechnologist. This is a full time permanent position on the night shift (12:30a-8:30a). My client offers excellent benefits and compensation and a generous shift differential. ASCP certification and strong experience in grossing is required. For more information please contact Pam Barker at 866-607-3542 or rel...@earthlink.net Thank You! Pam Barker President RELIA Solutions Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 Toll Free: (866)607-3542 FAX: (407)678-2788 E-mail: rel...@earthlink.net <mailto:rel...@earthlink.net> <<http://home.earthlink.net/~relia1>> www.myspace.com/pamatrelia www.twitter.com/pamatrelia ------------------------------ Message: 23 Date: Wed, 14 Oct 2009 12:34:59 -0400 From: Emily Sours <talulahg...@gmail.com> Subject: Re: [Histonet] unsubscribe, please To: susan bryant <sbry...@labpath.com>, histonet@lists.utsouthwestern.edu Message-ID: <b39794b0910140934v3417513fr82ad43a9ecad2...@mail.gmail.com> Content-Type: text/plain; charset=UTF-8 Good for you! You should probably FOLLOW THE LINK IN EVERY EMAIL JUST LIKE WE SAY EVERY TIME SOMEONE WRITES IN TO UNSUBSCRIBE. Emily ...the thrill of being close to that hidden knowledge. That's the way I feel when I read Nabokov. Encrypted within his words, encoded indecipherably, ambiguously, is the equivalent of the secret of lightning. Something akin to the secret code of higher human consciousness, the DNA, the genome of genius. -Ron Rosenbaum On Wed, Oct 14, 2009 at 11:24 AM, susan bryant <sbry...@labpath.com> wrote: > I wish to unsubscribe and return in 6 weeks. > > Thank you, > Susan E. Bryant > Knoxville Dermatopathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 24 Date: Wed, 14 Oct 2009 12:56:08 -0400 From: Peter Carroll <carro...@umdnj.edu> Subject: Re: [Histonet] unsubscribe, please To: Emily Sours <talulahg...@gmail.com> Cc: histonet@lists.utsouthwestern.edu, susan bryant <sbry...@labpath.com> Message-ID: <4ad602a8.6050...@umdnj.edu> Content-Type: text/plain; charset=UTF-8; format=flowed >FOLLOW THE LINK IN EVERY EMAIL JUST LIKE WE SAY EVERY TIME SOMEONE WRITES IN TO UNSUBSCRIBE Histonet Fun-Fact: Hits for the word "unsubscribe" in the archive: 1144 Hits for the word "eosin" in the archive: 801 Perhaps we should rename this list "Unsubscribenet" ;) ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 71, Issue 14 **************************************** This e-mail message (including any attachments) is for the sole use of the intended recipient(s) and may contain confidential and privileged information. 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