RE: [Histonet] FREEZY spray
Hey, Would you believe that putting the word Taupin into my outlook rules actually works. Now every obnoxious post from Taupin actually disappears into my Junk folder and I do not have to read the drivel. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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 Bernie Taupin Sent: Wednesday, 8 April 2009 3:38 PM To: Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] FREEZY spray Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq. From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:27:35 AM Subject: Re: [Histonet] FREEZY spray Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo..com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds.. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned.. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo..com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does
RE: [Histonet] FREEZY spray
Say thank you Uncle Kemlo!! Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: 08 April 2009 08:58 To: Bernie Taupin; Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: RE: [Histonet] FREEZY spray Hey, Would you believe that putting the word Taupin into my outlook rules actually works. Now every obnoxious post from Taupin actually disappears into my Junk folder and I do not have to read the drivel. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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 Bernie Taupin Sent: Wednesday, 8 April 2009 3:38 PM To: Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] FREEZY spray Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq. From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:27:35 AM Subject: Re: [Histonet] FREEZY spray Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo..com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care
Re: [Histonet] FREEZY spray
I see that Kemlo is an HistoNet charlatan like the rest of us. He's the first complain about off-topic notes to the list, then goes on to send several of his own. Thank you Uncle Kemlo! :-P Way to keep this ball rolling! From: Kemlo Rogerson kemlo.roger...@waht.swest.nhs.uk To: Tony Henwood antho...@chw.edu.au; Bernie Taupin bernietau...@ymail.com; Akemi Allison-Tacha akemiat3...@yahoo.com; Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 4:05:39 AM Subject: RE: [Histonet] FREEZY spray Say thank you Uncle Kemlo!! Kemlo Rogerson e-mail kemloroger...@nhs.net if not at work. DD 01934 647057 or extension 3311 Mob 07749 754194; Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: 08 April 2009 08:58 To: Bernie Taupin; Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: RE: [Histonet] FREEZY spray Hey, Would you believe that putting the word Taupin into my outlook rules actually works. Now every obnoxious post from Taupin actually disappears into my Junk folder and I do not have to read the drivel. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) 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 Bernie Taupin Sent: Wednesday, 8 April 2009 3:38 PM To: Akemi Allison-Tacha; Jennifer MacDonald; Ingles Claire Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Subject: Re: [Histonet] FREEZY spray Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq. From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:27:35 AM Subject: Re: [Histonet] FREEZY spray Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo..com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections
[Histonet] FREEZY spray
I have to agree with Jennifer on the subject of using freezing spray in cryostats containing potentially infectious agents. I have tried to promote the concept of good cryostat hygiene with everyone I train. When I started out in practice I was greeted by cryostats that have been left full of shavings. At our hospital we pathologists cut our own frozens, and it seemed to be the policy that the shavings were emptied when you could no longer close the door! One day I opened the door of the cryostat and the draft created by opening the door sent a small blizzard up and I literally inhaled shavings. From that moment on I demanded our trays be emptied and wiped clean after each use. It is tough to get some pathologists to comply as they think they are above menial cleaning tasks, and prefer there histoslaves to do there clean up. I believe these same culprits wait for their mothers to flush for them. Filthy cryostats also risk cross contamination of slides. If you drop your chuck it will come up covered in coconut. If you chunk out a precious piece of tissue in a freshly cleaned cryostat you may actually see it be able to re embed it. If you use freezing sprays in a cryostat snowstorm you will end up with a blizzard and risk not only inhalation but eye contact as well. If you must use freezing sprays, I suggest putting a dot on the chuck at 12:00, removing the chuck and spray the thing back to the ice age if you like. Return it to the chuck holder with the dot in the 12:00 position. Bring the chuck back a bit so you can start trimming over again without yet hitting the tissue in case there has been slight change in X-Y orientation. When you are done wipe it out and empty the tray. If you think this is over kill I can recommend a few polluted Beach's here in the New York metropolitan area to take your family this summer! Stephen Peters M.D. Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 Phone and fax 201 847 7600 www.pathologyinnovations.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] FREEZY spray
Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE: Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned. Regards,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ 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] FREEZY spray
I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the wayThis came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ Histonet mailing list histo...@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] FREEZY spray
Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE: Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned.. Regards,Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo..com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ 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] FREEZY spray
Sorry, again, I'm confused... are you responding to me? I'm not the original poster... I never stated I had trouble with cracking, because, well, I don't. I'm Bernie Taupin, the King of Cryomicrotomy, Esq. From: Akemi Allison-Tacha akemiat3...@yahoo.com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:27:35 AM Subject: Re: [Histonet] FREEZY spray Sorry, I too noticed that the text looked weird then I did a cut and paste. I keep all the Data Sheets I created in my files, and this was just a portion of the Notes from the original Data sheet. I thought it might be helpful because you stated you had difficulty with cracking. You did not mention using isopentane. Some people immerse the tissue straight into liquid nitrogen, which could cause freezing artifacts. Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Akemi Allison-Tacha akemiat3...@yahoo.com, Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 10:11 PM I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...? From: Akemi Allison-Tacha akemiat3...@yahoo..com To: Jennifer MacDonald jmacdon...@mtsac.edu; Ingles Claire cing...@uwhealth.org; Bernie Taupin bernietau...@ymail.com Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] FREEZY spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the MATSSE (pH3.4) 1-Step Trichrome Stain Kit (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way...This came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds.. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned.. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo..com --- On Tue, 4/7/09, Bernie Taupin bernietau...@ymail.com wrote: From: Bernie Taupin bernietau...@ymail.com Subject: Re: [Histonet] FREEZY spray To: Jennifer MacDonald jmacdon...@mtsac.edu, Ingles Claire cing...@uwhealth.org Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FREEZY spray
I was taught that freeze spray shouldn't be used - ever - on any tissue at all. We were told that it damages the tissue. Am I wrong?? We don't even use it when doing frozens - we have a different system now that extracts all the warmth and freezes the tissue completely so that we don't need the spray at all. Thanks for everyone's input on this! It will answer a lot of questions in our lab! Amy HT Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] FREEZY spray
Federal registry 1910.1030 “All procedures involving blood or other potential infectious materials shall be performed in such a manner as to minimize splashing, spraying, spattering, or generation of droplets of these substances.” NCCLS Document M29-T (now CLSI) “Frozen sections done on unfixed tissue pose a high risk because accidents are common. Freezing of tissue does not inactivate infectious agents. Freezing propellants under pressure should not be used for frozen sections as they may cause spattering of droplets of infectious material.” Based on the above statements, freezing spray should NOT be used in a cryostat with unfixed tissue. Jennifer MacDonald Senn, Amy R ars...@hsh.org Sent by: histonet-boun...@lists.utsouthwestern.edu 04/01/2009 08:25 AM To Histonet@lists.utsouthwestern.edu cc Subject [Histonet] FREEZY spray I was taught that freeze spray shouldn't be used - ever - on any tissue at all. We were told that it damages the tissue. Am I wrong?? We don't even use it when doing frozens - we have a different system now that extracts all the warmth and freezes the tissue completely so that we don't need the spray at all. Thanks for everyone's input on this! It will answer a lot of questions in our lab! Amy HT Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. 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
