[Histonet] Intestinal mucus -- Carnoy or what?
Hello! What is the best way to fix intestinal samples in order to preserve the mucus (and the embedded bacteria)? I was recommended Carnoy's, but are there any alternatives... perhaps without chloroform? With best regards, Mikael ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
Hi all Is anyone of you using a an alternative for the DAKO-ARK kit. In my opinion it is good, but rather expensive. Best regards, Joost Bruijntjes ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] embedding of insects in historesin (Technovit 7100)
Hallo, are there any experiences in embedding of insects in historesin (GMA or Glycolmethacrylat (Technovit 7100))? I have tried Technovit after fixing in Formaldehyd with poor results. The specimen tends to fail out of the block. There is no connection between the cuticula and the resin. Therefore it is not possible, to stretch the ribbon on the surface of water. And I believe there is some interaction between the chitin and the resin which makes trouble. The chitin is brittle after embedding in resin. I have tried different times in the embedding process, but without good results. Embedding single organs for example ovaries is without any difficulties . Obviously chitin makes the problem. Perhaps helps another fixing or some other pretreatment? Thanks for a little help ! J�rgen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] sectioning bee's heads and mites
___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Intestinal mucus -- Carnoy or what? (and: dry methanol??)
Oh, sorry, I forgot one additional question regarding Carnoy's: in the protocols I found, Carnoy's is prepared using DRY methanol. Sorry for my ignorance, but does this mean that methanol has to be specifically treated to remove any hygroscopic etc water? Note that this is specifically intended for preserving intestinal mucus, which I guess might be sensitive to water-containing fixatives (although I presume there can't be more water in ordinary methanol than introduced by the tissue sample itself). With best regards, Mikael On 21.11.2014 12:23, Mikael Niku wrote: Hello! What is the best way to fix intestinal samples in order to preserve the mucus (and the embedded bacteria)? I was recommended Carnoy's, but are there any alternatives... perhaps without chloroform? With best regards, Mikael ___ 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] embedding of insects in historesin (Technovit 7100)
First of all, this sounds like a Damien Laudier question. Damien, I hope you can help this guy. Secondly, do you have to use plastic embedding? I was successful with paraffin embedding. I was able to section various small insects and also bees. It takes some patience and soaking in mollifex or glycerin water but the chitin became less brittle and able to section without ripping out the internal organs. Andi G. From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of dr.ha...@gmx.net [dr.ha...@gmx.net] Sent: Friday, November 21, 2014 4:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] embedding of insects in historesin (Technovit 7100) Hallo, are there any experiences in embedding of insects in historesin (GMA or Glycolmethacrylat (Technovit 7100))? I have tried Technovit after fixing in Formaldehyd with poor results. The specimen tends to fail out of the block. There is no connection between the cuticula and the resin. Therefore it is not possible, to stretch the ribbon on the surface of water. And I believe there is some interaction between the chitin and the resin which makes trouble. The chitin is brittle after embedding in resin. I have tried different times in the embedding process, but without good results. Embedding single organs for example ovaries is without any difficulties . Obviously chitin makes the problem. Perhaps helps another fixing or some other pretreatment? Thanks for a little help ! J�rgen ___ 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] Thermo Scientific Richard-Allan Mark-it dyes
I already contacted them about this, but I thought that all of you might be faster: Would anyone happen to know if these dyes, if used to mark mouse bone/tissue around said bone at necropsy, would survive decalcification? We're trying to facilitate orientation of OA mouse knees during embedding Thanks and have a good weekend, Kathleen Roberts Principal Lab Technician Histopathology Lab Office of Translational Sciences Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (848) 445-1443 FAX (732) 445-6905 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] embedding of insects in historesin (Technovit 7100)
Hallo, Andi G. Thanks for your reply. Normally I use paraffine for embedding insects after dehydration and using N Butylalkohol with sucess. In my opinion it is absolutly necessary to avoid any rests of water and to cut cool. But this technique is limited to sections of 3-4 � and heating is unavoidable. I would be very grateful for a reply of Damien Laudier or of someone else, who could help me in this matter. J�rgen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Use of Polar Heat Pad inside Microwave Processor
I need a good, reliable company to buy from. This pad is not cheap. Thank you. Lin. Lin S. Bustamante, B.S., H.T.(ASCP) VIBS Histology Laboratory Supervisor College Of Veterinary Medicine Texas AM University College Station, Texas 77843-4458 Phone: (979) 845-3177 Fax: (979) 458-3499 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Use of Polar Heat Pad inside Microwave Processor
http://www.instructables.com/id/Microwave-Heating-Pad/ Like this? On Fri, Nov 21, 2014 at 12:21 PM, Bustamante, Lin lbustama...@cvm.tamu.edu wrote: I need a good, reliable company to buy from. This pad is not cheap. Thank you. Lin. Lin S. Bustamante, B.S., H.T.(ASCP) VIBS Histology Laboratory Supervisor College Of Veterinary Medicine Texas AM University College Station, Texas 77843-4458 Phone: (979) 845-3177 Fax: (979) 458-3499 ___ 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] Fast Green
Hey there, does anyone have a good method to de-stain Fast green? The slides need to be re-stained with HE. Thanks. Rebecca Berger, HT(ASCP)CM Research Technician Division of Orthopedic Surgery The Children's Hospital of Philadelphia Research Institute 3615 Civic Center Boulevard, ARC904 (Lab) Philadelphia, PA 19104 Phone: 267-425-2076 Email: berge...@email.chop.edumailto:berge...@email.chop.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Specimen numbering systems
Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] lab math: ihc dilution
To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Recycler
We have a CBG Biotech recycler. We recycle Americlear, xylene substitute. How does everyone dispose of the waste alcohol and waste paraffin? Thanks and have a great weekend! Tanya G. Abbott RT (CSMLS) Manager Technologist, Histology/Cytology St. Joseph Medical Center Reading, PA 19603-0316 ph 610-378-2635 fax 610-898-5871 email: tanyaabb...@catholichealth.net ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Specimen numbering systems
Numeric for the Specimen and Alpha for the Block. Joelle Weaver MAOM, HTL (ASCP) QIHC From: donna.wil...@baylorhealth.edu To: histonet@lists.utsouthwestern.edu Date: Fri, 21 Nov 2014 19:02:56 + Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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: Specimen numbering systems
Also, How are you specimens identified on your Surgical Requisition, 1,2,3 or A, B, C. Do you use the same identifiers in your LIS and Surgery. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Willis, Donna G. Sent: Friday, November 21, 2014 1:03 PM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonetd=AAIFAgc=qhent5lL-8Lans1hhN7NTGhSd0GBLfQfwUvzHj1D5tQr=holbn1kbGwnoiqK76TFBdjlG1OLqpvUtZAfF_4NbNVgm=6OI3m7sQR5qu-Enh6Z6bGbmilHJ2Y2lfwXlUfWRJIyks=EKDIAnC01YxcqQxvywzQK1ODb6WlOUVUt0GH3b63wKke= ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: lab math: ihc dilution
Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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] Specimen numbering systems
Same here. Brian D. Cooper, HT (ASCP)CM | Histology Supervisor Children's Hospital, Los Angeles -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joelle Weaver Sent: Friday, November 21, 2014 11:30 AM To: Willis, Donna G.; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Specimen numbering systems Numeric for the Specimen and Alpha for the Block. Joelle Weaver MAOM, HTL (ASCP) QIHC From: donna.wil...@baylorhealth.edu To: histonet@lists.utsouthwestern.edu Date: Fri, 21 Nov 2014 19:02:56 + Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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 - CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or legally privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of this original message. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Recycler
The waste alcohol is disposed of with the rest of our waste chemicals in a 55 gallon drum that picked is up by a licensed waste hauler. The paraffin is also picked up. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Abbott, Tanya Sent: Friday, November 21, 2014 12:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Recycler We have a CBG Biotech recycler. We recycle Americlear, xylene substitute. How does everyone dispose of the waste alcohol and waste paraffin? Thanks and have a great weekend! Tanya G. Abbott RT (CSMLS) Manager Technologist, Histology/Cytology St. Joseph Medical Center Reading, PA 19603-0316 ph 610-378-2635 fax 610-898-5871 email: tanyaabb...@catholichealth.net ___ 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: lab math: ihc dilution
Sheryl I will add another bit of information. That final antibody protein concentration is very low. Most of our antibodies (around 60 to 70%) will be in the range of 1 to 10 ug/ml. We do have some antibodies that can have some very low protein concentration though such as neurofilament. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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: lab math: ihc dilution
Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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
RE: [Histonet] RE: lab math: ihc dilution
Kathleen I have a final volume of 10 mls not 1 ml in the equation I wrote. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.commailto:l...@premierlab.com www.premierlab.comhttp://www.premierlab.com/ March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 From: Kathleen Jones [mailto:kjo...@upei.ca] Sent: Friday, November 21, 2014 12:50 PM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.commailto:l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.commailto:l...@premierlab.com www.premierlab.comhttp://www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ Histonet mailing list Histonet@lists.utsouthwestern.edumailto:Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edumailto: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] RE: lab math: ihc dilution
When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Jones Sent: Friday, November 21, 2014 11:50 AM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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
RE: [Histonet] RE: lab math: ihc dilution
You are right I got it wrong, see no matter how many time you run the math, you still come up with problems Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: Truscott, Tom [mailto:ttrus...@vetmed.wsu.edu] Sent: Friday, November 21, 2014 1:01 PM To: Kathleen Jones; Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: RE: [Histonet] RE: lab math: ihc dilution When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Jones Sent: Friday, November 21, 2014 11:50 AM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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] RE: Specimen numbering systems
We are currently using Numeric for the specimen (or part) and Alpha for the block. I don't like it; we frequently have 1Z blocks for large CA resections! I would like to change this going forward. 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) 972-1596 (860) 545-2204 Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Willis, Donna G. Sent: Friday, November 21, 2014 2:03 PM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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
[Histonet] RE: Specimen numbering systems
We do Alpha numeric so A1,A2 but our samples also get barcodes so we have a barcode ID that picks up whatever you designate it to and it is AAA001234 (designating a block) the slides then become AAA001234 0001, AAA001234 0002 etc. ties it back to the block and uniquely identified (called parent and children) The hitch here is if you had, say a colon it would be the (parent) and each block created would be a child and then technically the slides become the grandchildren. It works, but can be confusing at times - we're still working out the kinks. Turns out NCI and NIH use the system. Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cartun, Richard Sent: Friday, November 21, 2014 2:11 PM To: Willis, Donna G.; Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] RE: Specimen numbering systems We are currently using Numeric for the specimen (or part) and Alpha for the block. I don't like it; we frequently have 1Z blocks for large CA resections! I would like to change this going forward. 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) 972-1596 (860) 545-2204 Fax -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Willis, Donna G. Sent: Friday, November 21, 2014 2:03 PM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: lab math: ihc dilution
You are so right-that's why I always try to double check everything even my spellink. Tom T -Original Message- From: Elizabeth Chlipala [mailto:l...@premierlab.com] Sent: Friday, November 21, 2014 12:05 PM To: Truscott, Tom; 'histonet@lists.utsouthwestern.edu' (histonet@lists.utsouthwestern.edu) Subject: RE: [Histonet] RE: lab math: ihc dilution You are right I got it wrong, see no matter how many time you run the math, you still come up with problems Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: Truscott, Tom [mailto:ttrus...@vetmed.wsu.edu] Sent: Friday, November 21, 2014 1:01 PM To: Kathleen Jones; Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: RE: [Histonet] RE: lab math: ihc dilution When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Jones Sent: Friday, November 21, 2014 11:50 AM To: Sheryl Stephenson; histonet@lists.utsouthwestern.edu; Elizabeth Chlipala Subject: [Histonet] RE: lab math: ihc dilution Hi Liz, Wouldn't you dilute in 999.5ul of diluent, not .5. 1ml=1000ul Otherwise I got the same answer... Kathleen Kathleen Jones Research Technologist Pathology/Microbiology AVC - UPEI (902)213-2207 Elizabeth Chlipala l...@premierlab.com 21/11/2014 3:35 PM Sheryl Here is how I would approach the problem. We would use the basic mathematical equation of the following (in order for this to work you need to keep the units the same) V1 x C1 = V2 x C2 V1 - volume of stock solution needed - this is what you are solving for C1 - concentration of stock solution - you know this its 1.56 mg/ml or 1560 ug/ml (there are 1000 ug in one mg) V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed C2 - concentration of final solution - you know this its 0.8ug/ml V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml V1 = .0005 mls and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000. So in uls - V1 = 0.5 uls and .5 uls from there you can determine the titer which is your total volume divided by the volume of concentrated antibody 10/.0005 That would be a 1:20,000 dilution For a dilution such as this I would make up a stock solution of either 1:100 or 1:1000 and then dilute from there. Now its time for the rest of you to check my math, did I do this correctly? I went over it a few times, but you never know.. Have a GREAT Weekend! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl Sent: Friday, November 21, 2014 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab math: ihc dilution To all you math whiz out there, please help with this math dilution. If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use? Thanks, Sheryl ___ 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
Re: [Histonet] Specimen numbering systems
We use a numeric for the case number starting with the year 14-SR- and an alpha and number for blocs with the case. A1, A2, A3 so it looks like 14-SR- A1 and the next A block is A2 and so on with as many alpha designations as needed. Last week we had one surgical case with A through AA so it is somewhat flexible that way. Pam Marcum UAMS - Original Message - From: Donna G. Willis donna.wil...@baylorhealth.edu To: Histonet histonet@lists.utsouthwestern.edu Sent: Friday, November 21, 2014 1:02:56 PM Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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] Fast Green
Try 70% ETOH, leave the slides to soak for a good while. If that doesn't work after a couple of hours, try 1% HCl in 70% ETOH. Potassium permanganate followed by oxalic acid takes out just about anything, in my experience, but it's pretty harsh (1% aq potassium permanganate, 2% aq oxalic acid). Just like cleaning a stain on a shirt, you always want to start with the weakest solution first, and work your way up if necessary. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) On Fri, Nov 21, 2014 at 12:49 PM, Berger, Rebecca berge...@email.chop.edu wrote: Hey there, does anyone have a good method to de-stain Fast green? The slides need to be re-stained with HE. Thanks. Rebecca Berger, HT(ASCP)CM Research Technician Division of Orthopedic Surgery The Children's Hospital of Philadelphia Research Institute 3615 Civic Center Boulevard, ARC904 (Lab) Philadelphia, PA 19104 Phone: 267-425-2076 Email: berge...@email.chop.edumailto: berge...@email.chop.edu ___ 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] Reference to microtome micrometer thickness
Hello Histonetters, how will we verify the thickness of a paraffin section in a slide. Do we have a reference regarding on how to measure the thickness. I based the thickness of my section thru the mcirometer on the rotary microtome but one of our reveiwers does not believed that we are cutting them into 3 micron thickness or 4 micron thickness. Please if we have any reference, please share it to me. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: lab math: ihc dilution
Here's how I approach this problem. Convert all concentrations into the same units. I prefer to use whichever the final concentration unit is, so it looks like this: 1.56 mg/ml = 1560 ug/ml You have 1560 ug/ml and you need 0.8 ug/ml Divide 1560 by 0.8 = 1950. So your final dilution is 1:1950. Pipetting 1 ul of antibody in a dilution isn't very accurate, and using the above as 1 ul antibody in 1949 ul of diluent would yield you almost 2 ml of diluted antibody. Not a problem if you are doing batch staining where you might need 5 or 10 ml of diluted antibody. But if you are staining only 1 or 2 slides, you might not want to make that much. So follow Liz's advice about starting with a 1:10 dilution. Short division (move the decimal point one place to the left) puts the concentration at 156.0 ug/ml. 156 divided by 0.8 - 195 (You can see from above you could have just moved the final dilution decimal point one place to the left to get the same answer) With a 1:10 dilution of stock your final dilution would be 1:195 and that seems a bit more reasonable for fewer slides/total volume. Remember that 1:195 dilution is 1 part of antibody added to 194 parts of diluent. I would then multiply each side by the same number to get roughly how ever much diluted antibody you need. You can make *EXACTLY* however much you need, but that requires more complicated math, and I don't mind making just slightly over what I need to account for pipetting loss. Ex: If I need 100 ul/slide of diluted antibody and 4 slides to stain, I would multiply both sides by 3, since multiplying it by 2 won't give you quite enough : 1 x 3 = 3 (ul of 1:10 antibody) 194 x 3 = 582 (ul of diluent) Not sure if I helped or confused things. Teri Johnson, HT(ASCP)QIHC Manager Clinical Trial Testing Genoptix, Inc. SAN5, Rm. 2005 760.516.5954 (office) 760.516.6201 (fax) CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and contains information that is confidential and proprietary to Genoptix Medical Laboratory or its subsidiaries. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, immediately contact the sender by e-mail and destroy all copies of the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Help with muscle
Hello All, I hope I am not asking a dumb question. As I know that everyone on the histonet is very knowledgeable I was hoping to get some suggestions regarding processing tissue for neuromuscular junction staining. We are a neuro research lab and want to quantify neuromuscular junction on treated and non-treated rat gastrocnemius. We need the whole muscle for quantifications. The first problem we encountered when attempting to do this on PFA perfused and post fixed whole muscle was negative staining with SV2 antibody (presynaptic) but nicely stained alpha bungarotoxin (post synaptic) end plates. After a few attempts with no success we decided to freeze the whole muscle in dry ice and isopentane. First off we are having issues with the middle of the muscle freezing completely even after leaving it in solution for more than 10 min. Secondly while we do get nice staining with SV2 and bungarotoxin in some of the endplates we don’t see colocalization in most of the NMJ’s. One possibility that I was thinking could be causing this is that when the muscle is being picked up on the slide (cryosections) it is not laying completely flat on the slide (we tend to have to focus back and forth quite a bit) so we see the SV2/bungarotoxin near each other but not overlapping. One other thing I thought might be occurring is that when the muscle is being frozen it is retracting causing a shift in the presynaptic/postsynaptic NMJ. What is the best way to process the tissue, fixation or freezing? Any suggestions are greatly appreciated. Thanks, Leslie Leslie Garcia Senior Histologist Clive Svendsen Lab Board of Governors Regenerative Medicine Institute Cedars-Sinai Medical Center 8700 Beverly Blvd. AHSP 8405 Los Angeles, CA 90048 Phone - 310-248-8571 Web - http://www.cedars-sinai.edu/RMI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Specimen numbering systems
I am doing the same of Joelle Best Regards, Jamal M. Al Rowaihi Anatomic Pathology Supervisor | Al Borg Medical Laboratories | Mobile +966 503629832| j.rowa...@alborglaboratories.com Palestine St, Al Rajhi Building, P.O. Box 52817, Jeddah 21573, KSA| Phone: +966 12 670 0099 | Fax: +966 12 676 4984 | www.alborglaboratories.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joelle Weaver Sent: Friday, November 21, 2014 10:30 PM To: Willis, Donna G.; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Specimen numbering systems Numeric for the Specimen and Alpha for the Block. Joelle Weaver MAOM, HTL (ASCP) QIHC From: donna.wil...@baylorhealth.edu To: histonet@lists.utsouthwestern.edu Date: Fri, 21 Nov 2014 19:02:56 + Subject: [Histonet] Specimen numbering systems Can large facilities of more than 500 beds please let me know how they are numbering their Surgical specimens. Alpha for the Specimens and numeric for the Block or Numeric for the Specimen and Alpha for the Block. Thanks, Donna Willis, HT/HTL(ASCP) Anatomic Pathology Manager Baylor University Medical Center 3500 Gaston Ave|Dallas, Texas 75246 214-820-2465 office|214-725-6184 mobile BaylorScottandWhite.com ** This e-mail may contain confidential and/or privileged information. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden and possibly a violation of federal or state law and regulations. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. ___ 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
