[Histonet] re; cell blocks and frozen for IHC
No pretreatments for anything that is not formalin fixed. I think 95% ETOH for a fixative but but others may have a better idea than I. Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: cell blocks
I actually fixed the cells without the agar overnight, and left them in the agar overnight... Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: Tony Henwood [mailto:antho...@chw.edu.au] Sent: Tuesday, March 08, 2011 6:43 PM To: Sarah Goebel; histonet@lists.utsouthwestern.edu Subject: RE: cell blocks Sarah, It is important that the agar cell block is allowed to fix in formalin (10%NBF) before wrapping in paper, at least 2 hours. It will hold its shape better. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 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 sgoe...@mirnarx.com Sent: Wednesday, 9 March 2011 2:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cell blocks So...I am trying to make a cell block using 0.9% agar. I have done this in the past with no problems. It was at a different facility and I don't know if the agar is the same? I used bacto-agar this time, but last week someone tried to do my protocol with agarose and the boogers dried up and fell out of the block. The only other difference is we have clear rite xylene substitute in this processor where the other one I used had xylene. Tissues processed on the same cycle come out fine with the clear rite so I don't think that it the problem. When the cell blocks come out they are as thin as the papers I wrapped them in making it almost impossible to see and to subsequently cut. Any suggestions for making cell blocks without using the plasma/thrombin method or the insanely expensive histo-gel would be appreciated. Second, does anyone have a suggestion on how to repair the cells that are embedded in the tissue paper thick cell block? Thanks guys and gals! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: cell blocks
Sarah, The clear rite is the problem. We have it on our processor for routine paraffin processing and it works well. But if we use histogel or agarose, we make sure to use pure xylene ( not even recycled xylene). In past experiments, we have found that in the clear rite station the gel hardens, shrinks and discolors. Nancy -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@mirnarx.com Sent: Tuesday, March 08, 2011 9:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cell blocks So...I am trying to make a cell block using 0.9% agar. I have done this in the past with no problems. It was at a different facility and I don't know if the agar is the same? I used bacto-agar this time, but last week someone tried to do my protocol with agarose and the boogers dried up and fell out of the block. The only other difference is we have clear rite xylene substitute in this processor where the other one I used had xylene. Tissues processed on the same cycle come out fine with the clear rite so I don't think that it the problem. When the cell blocks come out they are as thin as the papers I wrapped them in making it almost impossible to see and to subsequently cut. Any suggestions for making cell blocks without using the plasma/thrombin method or the insanely expensive histo-gel would be appreciated. Second, does anyone have a suggestion on how to repair the cells that are embedded in the tissue paper thick cell block? Thanks guys and gals! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ 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: cell blocks
Sarah, It is important that the agar cell block is allowed to fix in formalin (10%NBF) before wrapping in paper, at least 2 hours. It will hold its shape better. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 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 sgoe...@mirnarx.com Sent: Wednesday, 9 March 2011 2:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cell blocks So...I am trying to make a cell block using 0.9% agar. I have done this in the past with no problems. It was at a different facility and I don't know if the agar is the same? I used bacto-agar this time, but last week someone tried to do my protocol with agarose and the boogers dried up and fell out of the block. The only other difference is we have clear rite xylene substitute in this processor where the other one I used had xylene. Tissues processed on the same cycle come out fine with the clear rite so I don't think that it the problem. When the cell blocks come out they are as thin as the papers I wrapped them in making it almost impossible to see and to subsequently cut. Any suggestions for making cell blocks without using the plasma/thrombin method or the insanely expensive histo-gel would be appreciated. Second, does anyone have a suggestion on how to repair the cells that are embedded in the tissue paper thick cell block? Thanks guys and gals! Sarah Goebel, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
