[Histonet] Cell block preparation (from cultured cells)
Hi everyone, Someone in the lab wishes to look at cultured cells in paraffin embedded cell blocks. These cells grow mostly in non-adherent clumps or cell bodies, which need to be preserved. Sections will be stained by IHC. I've been looking into the matter, and it seems there is not really a single straightforward method for this (as is always the case in histoland, apparently). I've been thinking to try fixing the cells in either 10% alcoholic formalin for 12-24 hours, or zinc-formalin. Then embed in agarose, fix some more with buffered formaldehyde and process. - What are your thoughts on the fixative of choice? - Should I process slowly, manually; or is a 'standard' ON processor protocol adequate (I don't know the exact timings, but I think it's a twelve hour protocol on a caroussel type processor). - Would Histogel have any added benefit over agarose? Thank you, Jonathan --- Jonathan Cremer Laboratory Technician TARGID - KU Leuven ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cell Block Preparation
We borrowed a wonderful cell block procedure from Windham Community Memorial Hospital here in CT several years ago. The specimen is collected fresh in sterile saline or RPMI and then centrifuged to concentrate the cells. Plasma and then thrombin are added to form a clot. The clot is then fixed in formalin and processed in our Histology Laboratory. I will send the procedure to anyone who is interested. Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax richard.car...@hhchealth.org From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of Ann Specian [thisis...@aol.com] Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann ___ 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 recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, or an employee or agent responsible for delivering the message to the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message, including any attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cell Block Preparation
This is how we do it now. In the old days, we used agar and to my mind, it is still the best way when you have scant material. - Spin in a conical tube and pour off - Melt an agar slant (we get TSA slant from micro) - Pour the agar into the conical tube and spin for 5 minutes - The agar will re-solidify and whatever sediment there is will be concentrated in the very tip of the cone - The agar will slide out of the centrifuge tube - Slice off the very tip and wrap in lens paper - Place the wrapped tip in a cassette and process as usual - Embed the specimen tip down and you are good to go... I still use this method today when I feel it necessary. Works great. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ann Specian Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cell Block Preparation
I wonder if this method could be used with the product Histogel. Has anyone tried it? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Friday, September 06, 2013 5:46 AM To: 'Ann Specian'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cell Block Preparation This is how we do it now. In the old days, we used agar and to my mind, it is still the best way when you have scant material. - Spin in a conical tube and pour off - Melt an agar slant (we get TSA slant from micro) - Pour the agar into the conical tube and spin for 5 minutes - The agar will re-solidify and whatever sediment there is will be concentrated in the very tip of the cone - The agar will slide out of the centrifuge tube - Slice off the very tip and wrap in lens paper - Place the wrapped tip in a cassette and process as usual - Embed the specimen tip down and you are good to go... I still use this method today when I feel it necessary. Works great. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ann Specian Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you. ___ 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] Cell Block Preparation
We recently switched most of our cell blocks from agar to Histogel, works great. We use small disposable embedding mold, put the specimen in the bottom and add the histogel about half way up the mold, let it harden, pop it out and put it in the cass. Also cuts much better than agar. Daniel Hewitt Histology Supervisor, HVS 412-749-7371 This email, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, or an agent responsible for delivering the message to the intended recipient, please contact the sender by reply e-mail and delete and destroy all copies of the original message, including attachments. Please note that any views or opinions presented in this e-mail are solely those of the author and do not necessarily represent those of Heritage Valley Health System. The integrity and security of this message cannot be guaranteed on the internet. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rathborne, Toni Sent: Friday, September 06, 2013 7:18 AM To: 'Tom McNemar'; 'Ann Specian'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cell Block Preparation I wonder if this method could be used with the product Histogel. Has anyone tried it? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Friday, September 06, 2013 5:46 AM To: 'Ann Specian'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cell Block Preparation This is how we do it now. In the old days, we used agar and to my mind, it is still the best way when you have scant material. - Spin in a conical tube and pour off - Melt an agar slant (we get TSA slant from micro) - Pour the agar into the conical tube and spin for 5 minutes - The agar will re-solidify and whatever sediment there is will be concentrated in the very tip of the cone - The agar will slide out of the centrifuge tube - Slice off the very tip and wrap in lens paper - Place the wrapped tip in a cassette and process as usual - Embed the specimen tip down and you are good to go... I still use this method today when I feel it necessary. Works great. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.org www.LMHealth.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ann Specian Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you. ___ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cell Block Preparation
This is a pretty good method for scant specimens. I have even used it for CSFs that have malignancy with success. http://www.jove.com/video/1316/cell-block-preparation-from-cytology-specimen-with-predominance CONFIDENTIALITY NOTICE: This e-mail message, including all attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. You may NOT use, disclose, copy or disseminate this information. If you are not the intended recipient, please contact the sender by reply e-mail immediately. Please destroy all copies of the original message and all attachments. Your cooperation is greatly appreciated. Columbus Regional Hospital 2400 East 17th Street Columbus, Indiana 47201___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cell Block Preparation
We have had mixed results in the past with cell blocks, but currently we are filtering the fluid through a biopsy bag and processing that. We seem to be retaining more specimen with that method (we have used sponges and lens paper in the past). Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General Hospital | An Affiliate of Commonwealth Health | mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 -Original Message- From: Ann Specian [mailto:thisis...@aol.com] Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
using celloidin RE: [Histonet] Cell Block Preparation
Our cytology department uses a technique that involves coating a centrifuge tube with Celloidin then spinning the sample in that tube. The cellodin is then scored and the bag of celloidin containing the cell button is taken out and wrapped in paper to be processed in a cassette. It is a tricky and a bit time consuming, but no material is lost in processing. We do up to 10 samples a day that way. Reference: J Clin Pathol. 1982 May; 35(5): 574-576. A celloidin bag for the histological preparation of cytologic material. G Bussolati Tim Morken Department of Pathology UC San Francisco Medical Center -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dessoye, Michael J Sent: Friday, September 06, 2013 9:39 AM To: Ann Specian; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cell Block Preparation We have had mixed results in the past with cell blocks, but currently we are filtering the fluid through a biopsy bag and processing that. We seem to be retaining more specimen with that method (we have used sponges and lens paper in the past). Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General Hospital | An Affiliate of Commonwealth Health | mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 -Original Message- From: Ann Specian [mailto:thisis...@aol.com] Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ 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] Cell Block Preparation
We spin the sample down, pour off supernatant. Resuspend in 5 drops thromboplastin and 4 drops human serum we receive from our phlebotomy lab. Let firm for 5 minutes and then fix in NBF. The agar suggestion probably works similarly if you can't get serum. On Sep 6, 2013 9:39 AM, Dessoye, Michael J mjdess...@commonwealthhealth.net wrote: We have had mixed results in the past with cell blocks, but currently we are filtering the fluid through a biopsy bag and processing that. We seem to be retaining more specimen with that method (we have used sponges and lens paper in the past). Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General Hospital | An Affiliate of Commonwealth Health | mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 570-552-1526 -Original Message- From: Ann Specian [mailto:thisis...@aol.com] Sent: Thursday, September 05, 2013 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cell Block Preparation I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 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 have received this email in error please notify the originator of the message. This footer also confirms that this email message has been scanned for the presence of computer viruses. Any views expressed in this message are those of the individual sender, except where the sender specifies and with authority, states them to be the views of Commonwealth Health. Scanning of this message and addition of this footer is performed by Websense Email Security software in conjunction with virus detection software. ___ 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] Cell Block Preparation
I am getting complaints in regard to insufficient cell blocks. We currently spin, pour off the supernatant, retrieve the sediment and process in lens paper. Does anyone have a more current technique which renders better cellularity? Also, do you know which renders a better cell block: a fresh specimen, a specimen fixed in Cytolyt or a specimen fixed in 10% NBF? Thanks, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
