From: | "Ian Birchall" |
Hi, after fixation with NBF I have performed H+E staining on the cells still adherent to the culture plate. Following fixation, wash with water, cover with haematoxylin (Harris) 5min, wash, blue (if necessary) wash, cover with eosin (3 min), wash and coverslip (on the culture plate) using aqueous mountant (Gurr, aquahere). You can also do immuno staining for cell surface markers on cultured cells without removing them from the plate. Regards, Ian Birchall, Melbourne. >>> histonet-request@lists.utsouthwestern.edu 10/8/03 3:00:01 AM >>> 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-admin@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. RE: Cell Culture Staining (C.M. van der Loos) 2. Re: Purchasing machines Grossing Work Station,Microtom etc. (Fred Underwood) 3. Re: Purchasing machines Grossing Work Station,Microtom etc. (Fred Underwood) 4. Nerve Tissue (LaTricia Faison) 5. Re: COX 1&2 (DDittus787@aol.com) 6. Hard Tissue Committee Meeting (Vicki Kalscheur) 7. Histotech needed (MGomez@ameripath.com) 8. For the Guiness Book Of Records (George Cole) --__--__-- Message: 1 Date: Tue, 07 Oct 2003 13:35:10 +0200 From: "C.M. van der Loos"To: histonet@lists.utsouthwestern.edu Cc: KJohnson@med.miami.edu Subject: [Histonet] RE: Cell Culture Staining Hi Kevin, There is a method described by Harold Kerstens to embed your cells into paraffin blocks. Spin the cells down in an Eppendorf tube, remove the supernatant and mix the cells gently with 1 ml warm (65C) 2% agarose solution. Centrifuge again to concentrate the cells. Embed in paraffin as usally. See paper by: Kerstens et al. JHC 48:709-718, 2000. For the next time try to get some collagen sponges from some cell culturing company. Spin the (unfixed!) cells down and wash 3x with PBS (remove proteins). Discard supernatant and resuspend cells in a small volume. Insert sponge and allow the cells to adhere to the collagen (15 min, RT). Fix sponge in buffered formalin and embed as usual. Good luck! Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >-----Original Message----- >From: Johnson, Kevin [mailto:KJohnson@med.miami.edu] >Sent: Friday, October 03, 2003 3:29 PM >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] Cell Culture Staining > >Greetings, all. > >A researcher just brought me (at Miller Time on a Friday!) two culture >flasks of terminally differentiated Sertoli cells and requested H&E >staining. Well. >Conventional H&E obviously is out, since polystyrene is not compatible >with xylene. Further restrictions: the cells cannot be scraped off, >they cannot be replated on glass coverslips, and for reasons of her >own, they cannot go back into the incubator pending an answer. They >now have been fixed in situ with 10% neutral buffered formalin. > >Does anyone have a protocol for staining these puppies? If not H&E >per se, then is there another staining method that might satisfy her? > >Thanks in advance, > >Kevin Johnson >University of Miami >Diabetes Research Institute --__--__-- Message: 2 Date: Tue, 07 Oct 2003 08:42:04 -0400 From: "Fred Underwood" To: , Subject: Re: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 3 Date: Tue, 07 Oct 2003 08:42:04 -0400 From: "Fred Underwood" To: , Subject: Re: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 4 Date: Tue, 07 Oct 2003 09:47:48 -0400 From: "LaTricia Faison" To: Subject: [Histonet] Nerve Tissue Does anybody know of a way to unthaw nerve that has been snap frozen to a = cork without doing damage to the nerve cells? It's hard to cross section = and remove the nerve from the cork when it is attached to it by snap = freezing. Thanks in advance for any advice. --__--__-- Message: 5 Date: Tue, 07 Oct 2003 10:25:11 -0400 From: DDittus787@aol.com To: Luis.Chiriboga@med.nyu.edu, dbpiontek@hsc.wvu.edu, histonet@pathology.swmed.edu Subject: Re: [Histonet] COX 1&2 you can try zymed as well, recently used both and they worked well. 1-800-874-4494 dana --__--__-- Message: 6 From: "Vicki Kalscheur" To: "Histonet Discussion" Date: Tue, 7 Oct 2003 11:13:19 -0500 Subject: [Histonet] Hard Tissue Committee Meeting This is a multi-part message in MIME format. ------=_NextPart_000_00A6_01C38CC4.03368850 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Dear members and interested individuals, Please join us on Saturday, October 18th, 2003 from 5:30 - 6:30 pm = in Louisville, KY as we gather at NSH's 29th Annual = Symposium/Convention. The room number will be posted at the Hard Tissue = Display Table. I look forward to seeing many of you!=20 Sincerely,=20 Vicki Kalscheur, Chairperson ------=_NextPart_000_00A6_01C38CC4.03368850 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Dear members and interested=20 individuals,Please join us on = Saturday,=20 October 18th, 2003 from 5:30 - 6:30 pm in Louisville, KY as we gather at = NSH's=20 29th Annual Symposium/Convention. The room number will be posted = at the=20 Hard Tissue Display Table. I look forward to seeing many of you! =Sincerely,Vicki Kalscheur,=20 Chairperson------=_NextPart_000_00A6_01C38CC4.03368850-- --__--__-- Message: 7 From: MGomez@ameripath.com To: Histonet@lists.utsouthwestern.edu Date: Tue, 7 Oct 2003 12:11:54 -0400 Subject: [Histonet] Histotech needed Good morning Histonetters: We are looking for a certified HT/HTL to work at our Histology Laboratory located in Sandy, Utah. If you know anyone interested please write or call at 801-256-0040 x205/x244 for immediate consideration. You may ask for Milton or Mike. We're located 20 minutes from Salt Lake City and 40 minutes from Park City. Thank you very much, Milton --__--__-- Message: 8 From: "George Cole"To: Date: Tue, 7 Oct 2003 09:11:01 -0700 Subject: [Histonet] For the Guiness Book Of Records This is a multi-part message in MIME format. ------=_NextPart_000_000F_01C38CB2.EE7ECAA0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit We have a place in the Guiness Book Of Records----Egad!! Who would ever think there would be so many histotechs named Markus F. Meyenhofer. ------=_NextPart_000_000F_01C38CB2.EE7ECAA0 Content-Type: text/html; charset="us-ascii" Content-Transfer-Encoding: quoted-printable ------=_NextPart_000_000F_01C38CB2.EE7ECAA0-- --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonetWe have a place in the Guiness Book Of Records----Egad!! Who would ever = think there would be so many histotechs = named
Markus = F. Meyenhofer.