RE: [Histonet] RE: time off
Time off is one thing but holidays are another. Holidays need to be rotated. Seniority should apply to things like choice of hours but not holidays. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, January 04, 2012 8:47 AM To: Andrea; Toni Rathborne Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: time off I've witnessed that granting time by seniority lets you open to abuse. I know of an employee with high seniority who takes the whole week after Christmas every year. Our policy allows only one person off per day per shift. So then no one else ever can take off at Christmastime. Her peers complain to management about it, but they won't say anything to her. Just my two cents. Andrea abeharry...@gmail.com 1/3/2012 5:40 PM We also have a policy on the number of people that can be off during peak times. Our vacation schedule runs from June to June of the following year. Staff have up until a deadline of January 31 to put in their request for the next vacation year, whether it is one day or weeks. The requests handed in by this time period are granted based on seniority. ( in our institution they figure it's one of the only perks to being a senior!) After the January 31 deadline it is first come first serve. All vacation requests must be submitted on a vacation request form and time stamped when handed in. This way if two people ask for the same day the person who handed it in with an earlier time stamp is granted the time off. All of this is written in our scheduling guidelines. It seems to work pretty well. On 2012-01-03, at 4:32 PM, Rathborne, Toni trathbo...@somerset-healthcare.com wrote: We have a Laboratory policy which states that holidays will be rotated. There is also a section which gives a limit to the amount of time an employee can have off during peak vacation time. For example, our staff can only have a maximum of two weeks off during the peak summer time, and no more that 2 days off during the last two weeks of December. I personally have no problem with staff requesting time off early in the year, but I do ask that they discuss with their coworkers before giving me the request. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amber McKenzie Sent: Tuesday, January 03, 2012 4:10 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] time off Those of you who are supervisors, how do you handle your co-workers asking for time off? I have 2 employees that have asked off already (jan 3rd) for every day they want off for the entire year! Do you grant them the days off since no one else has asked off yet, or tell them it's not fair to continuously get off around every holiday by asking off 5 - 12 months in advance? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Be sure to visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for the most up-to-date news, event listings, health information and more. ___ 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 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately
[Histonet] Histological mordant
Hi all, I was going to prepare a solution 1% in distlled water of dodecaphosphomolybdic acid as histological mordant for Mallory staining. I noticed that it was not completely soluble in water, as the Chemistry says, but in the yellowish solution there was a thin precipitate quite heavy. So I have to filter the solution. I wonder if there will be a leak as mordant or it will be necessary to lengthen the processing time. Any experience about? Thank you in advance. Kind Regards, Massimo Tosi ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] NCCI policy on IHC billing
Sally, This is a little different from what I understood yesterday. I thought that it was stated you could only bill once per part, such as an S-100 on part A,B,C would be fine, but not on A1,A3, and A5. I did not read anything about only being allowed to order one antibody. I thought panels were understood and completely acceptable as long as you were only billing for each individual stain once per part. Also, I understood the part about cocktail stains now being billed as one charge instead of multiple. Do you have a direct link to what you are stating? I mentioned it to our powers that be yesterday and they did not seem too concerned since they had heard absolutely nothing about it from our billing company. I would like to have some concrete proof before I go back to them with more bad news. Thanks, Sheila, HT (ASCP) KDL Pathology Knoxville, TN -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sally Price Sent: Thursday, January 05, 2012 12:17 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] NCCI policy on IHC billing Histonetters: I waited a few days to see how others might weigh in after this information was posted. Call me crazy, but I expected quite a bit more reaction from our community. How is it that such a signifcant change in how IHC testing may be conducted and will be paid for in the future can produce so little response? The way this new policy is stated, it looks pretty straightfoward: one antibody (IHC procedure) per specimen; so, when it's necessary to use a battery of IHC stains to determine the origin of an undifferentiated neoplasm, the lab can only bill for one procedure. How could such an approach be possible? And what about multi-antibody procedures, which are usually more cost effective than single-antibody procedures? Come on folks, this is a big deal becuase IHC staining is essential to to the practice of anatomic pathology and provides a lot of us with our livelihood. I know I'm not alone in thinking that the CMS needs to know that this new policy is completely impractical and must be changed. Sure, there's some unnecessary IHC procedures being performed, but this isn't the way limit the problem. Sally -- Message: 6 Date: Fri, 30 Dec 2011 12:33:17 -0600 From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] NCCI policy update To: 'histonet@lists.utsouthwestern.edu' Is everyone aware that beginning 1/1/12, we can no longer bill for each block regarding IHC billing, only one unit of billing for each part type no matter how many blocks are stained? Also IHC cocktail stains, such as PIN4 must now be billed as one unit even though multiple antibodies are reported out. Kind of a surprising reversal of the policy set in motion 10/1/2009. SPECIMEN becomes the unit of service rather than block(s) for IHC codes 88342, 88360, and 88361. Happy New Year to everyone out there. May 2012 find you happiness and health! Dorothy Webb, HT Regions Histology TS 651-254-2962 ___ 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: Using Nail Polish on coverslip?
I'll seal the coverslip around all the edges and let it dry in the hood for up to half an hour (because of the smell). I use a clear color that doesn't fluoresce under the miscroscope and also allows the slide to be stored at low temp (-20) without the nail polish seal cracking. Also be sure the polish is toluene- and benzene-free. We've discovered Revlon #771 Clear fits these parameters and works very well. Cheaper nail polishes seem to crack or flake off at cold temps. Regards, Merced -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kasai, Miki (NIH/NCI) [E] Sent: Wednesday, January 04, 2012 11:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Using Nail Polish on coverslip? We are currently starting up some IHC on frozen tissue sections. After staining with different fluorescent antibodies, we end with applying DAPI w/Prolong gold and then coverslipping. We would like to seal the coverslip so that we can keep the slides longer. Any suggestions on where and how best to apply the nail polish for a permanent fix on the coverslips? Thanks, ___ 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] Cutting speed
Theresa Although I agree with the messages on not trading quality for quantity we ask our new grads to meet the following within the first 3 months. Embedding 1 min/ block Paring and cutting 2 min/block (average) Some blocks are move difficult or require more levels and will take longer and some will take less time. Can you tell me if you received any numerical responses this will give me an idea if our requirement is out of line? Thanks and good luck! Sharon Genest Anatomic Pathology Process Improvement Saskatoon Health Region 306-655-8242 sharon.gen...@saskatoonhealthregion.ca ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] NCCI policy on IHC billing
From what we understand from the new policy the cocktailed antibodies, such as PIN4, are not to be charged separately. Martha Ward, MT (ASCP) QIHC Manager, Molecular Diagnostics Lab Dept. of Pathology Wake Forest University Baptist Medical Center Winston-Salem, NC 27157 336-716-2104 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rae Staskiewicz Sent: Thursday, January 05, 2012 9:00 AM To: 'Clare Thornton'; 'Sally Price'; histonet@lists.utsouthwestern.edu Cc: Dana Spears Subject: RE: [Histonet] NCCI policy on IHC billing Clair, I agree that the cocktailed antibodies would be charged as a single test, however, with the dual and triple stains, since these are technically done with separate procedures, would not each antibody be able to be charged? This is what we are thinking. How are others interpreting this Rae Staskiewicz -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Clare Thornton Sent: Thursday, January 05, 2012 5:35 AM To: Sally Price; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] NCCI policy on IHC billing Sally, The way I read it, I don't think it's one antibody per specimen; I think what they are saying is that if you are running one antibody on multiple blocks from one specimen, you can only bill once for that antibody. I think if you are running a panel of antibodies one one block from a specimen, you can still charge for each antibody. However, I completely agree with you. This change came as a big surprise for our lab, until Dorothy posted it on Histonet we had no idea. How can that be? (And thanks, Dorothy!) The day that post came out we happened to be running 20 pankeratin/p63 double stain slides, all on multiple blocks from one specimen. The two antibodies from that double stain are applied at different times (to the same slide) and we use two different detections. So had we ran them after the new year, we would've been able to charge only once for those 20 slides, never mind 20 tests being used from each antibody and 20 tests being taken from two different detection kits. And the tech time put into cutting those 20 slides! We have several double and triple stains that we run on a daily basis, and only one component from the triple stain is a cocktail; the rest are separate antibodies being applied to the same slide using two different detections. I'm not sure exactly what our lab is going to do about it, but somehow this change should have been made aware to everyone well before it happened. There are going to be labs who are no longer in compliance, and we all know what that means.. Clare J. Thornton, HTL(ASCP) Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sally Price [sprice2...@gmail.com] Sent: Thursday, January 05, 2012 12:16 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] NCCI policy on IHC billing Histonetters: I waited a few days to see how others might weigh in after this information was posted. Call me crazy, but I expected quite a bit more reaction from our community. How is it that such a signifcant change in how IHC testing may be conducted and will be paid for in the future can produce so little response? The way this new policy is stated, it looks pretty straightfoward: one antibody (IHC procedure) per specimen; so, when it's necessary to use a battery of IHC stains to determine the origin of an undifferentiated neoplasm, the lab can only bill for one procedure. How could such an approach be possible? And what about multi-antibody procedures, which are usually more cost effective than single-antibody procedures? Come on folks, this is a big deal becuase IHC staining is essential to to the practice of anatomic pathology and provides a lot of us with our livelihood. I know I'm not alone in thinking that the CMS needs to know that this new policy is completely impractical and must be changed. Sure, there's some unnecessary IHC procedures being performed, but this isn't the way limit the problem. Sally -- Message: 6 Date: Fri, 30 Dec 2011 12:33:17 -0600 From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] NCCI policy update To: 'histonet@lists.utsouthwestern.edu' Is everyone aware that beginning 1/1/12, we can no longer bill for each block regarding IHC billing, only one unit of billing for each part type no matter how many blocks are stained? Also IHC cocktail stains, such as PIN4 must now be billed as one unit even though multiple antibodies are reported out. Kind of a surprising reversal of the policy set in motion 10/1/2009. SPECIMEN becomes the unit of
[Histonet] RE: decal solution for molecular studies
To clarify, the Formical we use is a formic acid/EDTA solution. Clare J. Thornton, HTL(ASCP), QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Clare Thornton Sent: Thursday, January 05, 2012 9:12 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] decal solution for molecular studies Does anyone know of a decal solution that enables molecular studies (Her 2 FISH, EGFR by PCR) to be performed later? We use Formical for our decal solution. Her 2 FISH works about 50% of the time, EGFR almost never. We are looking at having to cut undecalcified bone today for both these studies, and we are not equipped to cut undecalcified bone. Any suggestions? Clare J. Thornton, HTL(ASCP), QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com ___ 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] Amazing
I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Amazing
If you are talking about a regressive HE stain then perhaps it was 1% HCL acid in 70% alcohol -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart Sent: Thursday, January 05, 2012 11:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Amazing I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Amazing
You can use either acetic or hydrochloric, both will do. The strength is 1% BUT it does not matter. As a matter of fact the weaker the solution the better. Remember that the acid solution (that can be made with 7ethanolanol even better) is used to differentiate progressivesive hematoxylin, like Harris, and the weaker it is the more control you have with the differentiation. A weak solution allows you to dip the slides several times until you obtain the desired reddish hue on the sections that signals when the differentiation is completed. If the solution is too strong you will have to take the slides often precipitouslyusly and some sections will be have a weak nuclear staining. So, use acetic at 1% in 70% ethanol That is what I used to prepare. René J. --- On Thu, 1/5/12, Sarah Dysart sdys...@mirnarx.com wrote: From: Sarah Dysart sdys...@mirnarx.com Subject: [Histonet] Amazing To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Thursday, January 5, 2012, 10:59 AM I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Amazing
I still use that. It is 70% with 5mL of hydrochloric acid if making 1L. Lynn Burton Lab Assoc I Animal Disease Lab Galesburg, Il 309-344-2451 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart [sdys...@mirnarx.com] Sent: Thursday, January 05, 2012 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Amazing I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Amazing
We use .5% acid alcohol ( hydrochloric) with Harris hemo. No problems. Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL 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 Rene J Buesa Sent: Thursday, January 05, 2012 10:17 AM To: histonet@lists.utsouthwestern.edu; Sarah Dysart Subject: Re: [Histonet] Amazing You can use either acetic or hydrochloric, both will do. The strength is 1% BUT it does not matter. As a matter of fact the weaker the solution the better. Remember that the acid solution (that can be made with 7ethanolanol even better) is used to differentiate progressivesive hematoxylin, like Harris, and the weaker it is the more control you have with the differentiation. A weak solution allows you to dip the slides several times until you obtain the desired reddish hue on the sections that signals when the differentiation is completed. If the solution is too strong you will have to take the slides often precipitouslyusly and some sections will be have a weak nuclear staining. So, use acetic at 1% in 70% ethanol That is what I used to prepare. René J. --- On Thu, 1/5/12, Sarah Dysart sdys...@mirnarx.com wrote: From: Sarah Dysart sdys...@mirnarx.com Subject: [Histonet] Amazing To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Thursday, January 5, 2012, 10:59 AM I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Look at me...
I'm just full of questions today!! This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo. I do all the blocking including Fc receptors...still junk. The clone I have been using is, from abcam (ab2302). I don't see the specific clone name listed. I am staining human xenografts raised in mouse. I get a whole lot of background staining making it very hard to find the positive staining. The recommended dilution is about 1:30, but I have diluted all the way up to 1:500. At the higher dilution no positive staining or background is observed. Does anyone know of a good Caspase3 antibody, preferably mouse monoclonal? All the rabbit polyclonal antibodies are difficult to stain on these xenografts. Thanks again =) Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Nail Polish sealant
You wrote: We are currently starting up some IHC on frozen tissue sections. After staining with different fluorescent antibodies, we end with applying DAPI w/Prolong gold and then coverslipping. We would like to seal the coverslip so that we can keep the slides longer. Any suggestions on where and how best to apply the nail polish for a permanent fix on the coverslips? * Prolong Gold antifade reagent is a hard seal to begin with. We only seal ends but not the sides of cover glass. Our coverslips go right to edge of slides e.g. 25 X 30, 25 x 40, or 25 X 50.We don't like having nail polish slop over the edges onto back of slides but one could seal all sides of coverslip if careful. We buy the thinner top or base coat nail polish, but in general, prefer to use thinned mounting media rather than nail polish. There can be some issues here.If you are trying to view GFP or RFP (red fluorescence protein) labeled cells or tissue components, you should not seal the coverslip with nail polish since the alcohol in nail polish leaches under the coverslip and causes GFP/RFP to fade. This fact was published in Science. We found that dumping out cheap clear nail polish from bottle, rinsing away the residue with acetone, and then adding permanent mounting media and thinning that with toluene to the consistency of top coat nail polish works best. Toluene or xylene based sealants cannot leach under the cover glass since these solvents are NOT miscible with water in the PBS.Thinned mounting media is better sealant for GFP purposes (no fading) and also works for IF stained sections (perfect seal). We love the little brush in the nail polish bottle for application. Thicker clear nail polish (for non GFP studies) or IF stained sections is messy during application so we buy the cheapest top coat polish we can find at Walmart. DAPI in the Prolong Gold will cause an uneven staining gradient so that some of the nuclei in the center of a section are not stained as brightly as the nuclei on the outer edges of a stained section. The cause is not getting enough thicker Prolong Gold/DAPI over the section or not having just the right amount of buffer on the section to permit a good flow of this wonderful mounting media over the section.We now complete all IF staining then stain with a DAPI solution before cover slipping with Prolong Gold. You can buy ready to use DAPI solutions from Pierce or Biogenex, or make up the solution in house. You can find the recipe at IHCworld website or simply Google. We do NOT store our IF stained slides in the cold, but in a folder at RT in a dark drawer before viewing on the day after staining to reduce any movement/flow under the coverslip. Fluorophores can and will eventually fade. I do not recall any studies saying storing IF stained slides in the cold reduces fading but we never have space to do cold storage anyway and store slides at RT.The new fluorophores (Alexas and Dylights) remain stable over a longer time even for several weeks compared to fluorescein derivatives e.g. FITC TRITC. Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Look at me...
If you are looking to stain cleaved caspase 3 there is a better antibody from cell signaling it works in multiple species, it's a bit pricey but I have found that it works the best out of the few that I have tried. We have used it mouse xenografts before without any issue. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 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 Sarah Dysart Sent: Thursday, January 05, 2012 10:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Look at me... I'm just full of questions today!! This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo. I do all the blocking including Fc receptors...still junk. The clone I have been using is, from abcam (ab2302). I don't see the specific clone name listed. I am staining human xenografts raised in mouse. I get a whole lot of background staining making it very hard to find the positive staining. The recommended dilution is about 1:30, but I have diluted all the way up to 1:500. At the higher dilution no positive staining or background is observed. Does anyone know of a good Caspase3 antibody, preferably mouse monoclonal? All the rabbit polyclonal antibodies are difficult to stain on these xenografts. Thanks again =) Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Elastichrome paper, and troubleshooting
Does anyone have a pdf (or could fax me) the original paper for Elastichrome stain: Richardson, L. Combination Elastic Trichrome Stain, Laboratory Medicine, 6.1, 1975 We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past. I do have another paper in the same vein, A combination verhoeff's elastic and masson's trichrom stain for routine histology, O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible. Our problem is that the elastic stain washes out during or after the trichrome. Procedure: Bouins, 56C one hour Wiegerts hematolylin, 3min Verhoffs elastic stain, 15 min 2% ferric chloride differentiation (so far so-good) Gomori's trichrome Blue, 15 min 0.2% glacial acetic acid, 1 min Wash dH20, dehydrate, clear, mount Results: trichrome looks great, no elastin stain. I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions. Thanks for any help. Tim Morken Supervisor, Histology, IPOX UC San Francisco Medical Center Box 1656 1600 Divisidero St, B217 San Francisco, CA 94115 USA 415.514.6042 (office) 415.885.7409 Fax tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Elastichrome paper, and troubleshooting
Tim We do this stain all of the time, we never used the original reference we just made up one on our own. I'll send our SOP in a different e-mail. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 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 Morken, Timothy Sent: Thursday, January 05, 2012 10:45 AM To: Histonet Subject: [Histonet] Elastichrome paper, and troubleshooting Does anyone have a pdf (or could fax me) the original paper for Elastichrome stain: Richardson, L. Combination Elastic Trichrome Stain, Laboratory Medicine, 6.1, 1975 We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past. I do have another paper in the same vein, A combination verhoeff's elastic and masson's trichrom stain for routine histology, O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible. Our problem is that the elastic stain washes out during or after the trichrome. Procedure: Bouins, 56C one hour Wiegerts hematolylin, 3min Verhoffs elastic stain, 15 min 2% ferric chloride differentiation (so far so-good) Gomori's trichrome Blue, 15 min 0.2% glacial acetic acid, 1 min Wash dH20, dehydrate, clear, mount Results: trichrome looks great, no elastin stain. I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions. Thanks for any help. Tim Morken Supervisor, Histology, IPOX UC San Francisco Medical Center Box 1656 1600 Divisidero St, B217 San Francisco, CA 94115 USA 415.514.6042 (office) 415.885.7409 Fax tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org ___ 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: Look at me...
I'll second that from our experience - Cell Signaling cleaved caspase-3 antibody works well on xenografts Brett Brett M. Connolly, Ph.D. Imaging Research Fellow Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_conno...@merck.com T- 215-652-2501 F- 215-993-6803 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Thursday, January 05, 2012 12:36 PM To: 'Sarah Dysart'; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Look at me... If you are looking to stain cleaved caspase 3 there is a better antibody from cell signaling it works in multiple species, it's a bit pricey but I have found that it works the best out of the few that I have tried. We have used it mouse xenografts before without any issue. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 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 Sarah Dysart Sent: Thursday, January 05, 2012 10:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Look at me... I'm just full of questions today!! This one is IHC...I have been trying to optimize a Caspase3 stain for several months now and it is still just chalked full of background gradoo. I do all the blocking including Fc receptors...still junk. The clone I have been using is, from abcam (ab2302). I don't see the specific clone name listed. I am staining human xenografts raised in mouse. I get a whole lot of background staining making it very hard to find the positive staining. The recommended dilution is about 1:30, but I have diluted all the way up to 1:500. At the higher dilution no positive staining or background is observed. Does anyone know of a good Caspase3 antibody, preferably mouse monoclonal? All the rabbit polyclonal antibodies are difficult to stain on these xenografts. Thanks again =) Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE:
Sarah, I would use the 1% Acetic Acid in 70% alcohol, for a couple of reasons: your docs are used to looking at the stain with the Clarifier, and acetic is the ingredient in the Clarifier 2; also you probably will not have to change your staining times by too much if you use acetic. You could probably even dilute it out to 50% alcohol. HCL works too fast, especially since I have students. Toysha N. Mayer, MBA, HT (ASCP) Instructor Program in Histotechnology School of Health Professions MD Anderson Cancer Center (713) 563-3481 tnma...@mdanderson.org -- Message: 2 Date: Thu, 5 Jan 2012 15:59:35 + From: Sarah Dysart sdys...@mirnarx.com Subject: [Histonet] Amazing To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: 8a70a9b2ecdd084dacfe6c59fcf86d509c9...@sn2prd0702mb098.namprd07.prod.outlook.com Content-Type: text/plain; charset=us-ascii I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Elastichrome Paper
Tim, I do not have the paper, but a few years ago I worked the kinks out of that stain. Your acetic acid is too long. Try for the amount of time that you usually do for a one step. A few dips would be a good start. Then rinse in dH2O, not wash. Also, I do not remember using the Weigert's and Verhoeff's solution. No need. Just the Bouins mordant, VVG and did a trichrome as a counter stain. Also, do not differentiate as much with the Ferric. Under differentiation will compensate for the other reagents. How long do you usually leave the slides in one step when doing a regular one? Those would be good starting points. Toysha N. Mayer, MBA, HT (ASCP) Instructor Program in Histotechnology School of Health Professions MD Anderson Cancer Center (713) 563-3481 tnma...@mdanderson.org -- Message: 10 Date: Thu, 5 Jan 2012 09:45:25 -0800 From: Morken, Timothy timothy.mor...@ucsfmedctr.org Subject: [Histonet] Elastichrome paper, and troubleshooting To: Histonet histonet@lists.utsouthwestern.edu Message-ID: 8d7c2d242dbd45498006b21122072bf89f5ee...@mcinfrwem003.ucsfmedicalcenter.org Content-Type: text/plain; charset=us-ascii Does anyone have a pdf (or could fax me) the original paper for Elastichrome stain: Richardson, L. Combination Elastic Trichrome Stain, Laboratory Medicine, 6.1, 1975 We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past. I do have another paper in the same vein, A combination verhoeff's elastic and masson's trichrom stain for routine histology, O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible. Our problem is that the elastic stain washes out during or after the trichrome. Procedure: Bouins, 56C one hour Wiegerts hematolylin, 3min Verhoffs elastic stain, 15 min 2% ferric chloride differentiation (so far so-good) Gomori's trichrome Blue, 15 min 0.2% glacial acetic acid, 1 min Wash dH20, dehydrate, clear, mount Results: trichrome looks great, no elastin stain. I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions. Thanks for any help. Tim Morken Supervisor, Histology, IPOX UC San Francisco Medical Center Box 1656 1600 Divisidero St, B217 San Francisco, CA 94115 USA 415.514.6042 (office) 415.885.7409 Fax tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Respirators and Routine Histology
Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Clearing NBT/BCIP precipitate from wax sections
Hello, I have some paraffin sections that I've used for immunohistochemistry (using an alkaline phosphatase-conjugated secondary antibody and NBT/BCIP as a substrate), and coverslipped with an aqueous mounting medium (Fluoromount G). I would now like to de-coverslip and re-stain these sections with a different histochemical protocol (i.e. Masson's trichrome or HE). Does anybody know if there is a way to clear away the colour reaction precipitate from my previous IHC? If, after decoverslipping, I dehydrate and then rehydrate the sections, will this do the job? Any advice would be greatly appreciated. Thanks very much, Andrew ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] saffron vs. safran du gatinais
Hello histonet, I have a protocol that calls for safran du gatinais and am experiencing some serious sticker shock. Can I substitute saffron in my solution or am I asking for trouble? Many thanks, Beth Beth Villarreal Scientist I Novartis Institutes for BioMedical Research, Inc. 300 Technology Square Cambridge, MA 02139 USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Respirators and Routine Histology
No, not here. We don't wear anything at all. We have excellent air flow, and we wear the badges that test for exposure to xylene and formalin annually which I think is a CAP requirement. Our results have been better than acceptable every year. Lisa Brenner HTL (ASCP) Histology Technical Consultant Holland Hospital phone: (616)394-3184 li...@hollandhospital.org Amy Self as...@georgetownhospitalsystem.org 1/5/2012 3:09 PM Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: The information contained in this e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information, or Protected Health Information as such term is defined under the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Any unauthorized review, use, disclosure, copying or distribution is prohibited and may be unlawful. If you believe you have received this e-mail in error, please contact the sender by reply e-mail and delete all copies of the original message, including attachments. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Respirators and Routine Histology
Nothing at all??? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lisa Brenner Sent: Thursday, January 05, 2012 3:22 PM To: Amy Self; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Respirators and Routine Histology No, not here. We don't wear anything at all. We have excellent air flow, and we wear the badges that test for exposure to xylene and formalin annually which I think is a CAP requirement. Our results have been better than acceptable every year. Lisa Brenner HTL (ASCP) Histology Technical Consultant Holland Hospital phone: (616)394-3184 li...@hollandhospital.org Amy Self as...@georgetownhospitalsystem.org 1/5/2012 3:09 PM Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: The information contained in this e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information, or Protected Health Information as such term is defined under the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Any unauthorized review, use, disclosure, copying or distribution is prohibited and may be unlawful. If you believe you have received this e-mail in error, please contact the sender by reply e-mail and delete all copies of the original message, including 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] Respirators and Routine Histology
They must be in a liberal part of Michigan. - Original Message - From: Linda Blazek lbla...@digestivespecialists.com To: Lisa Brenner li...@hollandhospital.org, Amy Self as...@georgetownhospitalsystem.org, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Thursday, January 5, 2012 2:30:20 PM Subject: RE: [Histonet] Respirators and Routine Histology Nothing at all??? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lisa Brenner Sent: Thursday, January 05, 2012 3:22 PM To: Amy Self; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Respirators and Routine Histology No, not here. We don't wear anything at all. We have excellent air flow, and we wear the badges that test for exposure to xylene and formalin annually which I think is a CAP requirement. Our results have been better than acceptable every year. Lisa Brenner HTL (ASCP) Histology Technical Consultant Holland Hospital phone: (616)394-3184 li...@hollandhospital.org Amy Self as...@georgetownhospitalsystem.org 1/5/2012 3:09 PM Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: The information contained in this e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information, or Protected Health Information as such term is defined under the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Any unauthorized review, use, disclosure, copying or distribution is prohibited and may be unlawful. If you believe you have received this e-mail in error, please contact the sender by reply e-mail and delete all copies of the original message, including 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] saffron vs. safran du gatinais
Hi Beth, Saffron and safran du gatinais both refer to the dried stigma of the saffron crocus. They are both saffron. I use and alcoholic extract of Saffron to stain collagen in Movat stains. I've tried saffron from multiple sources and price definitely does not always correlate with quality. I gave up on Sigma-Aldrich, for example. The best saffron has a deep red color and is highly aromatic. There are possible workarounds for the lower quality material--like use more saffron, or stain longer, but best to use the best. What stain are you doing and what solution are you making with the saffron? Jerry Ricks Research Scientist University of Washington Department of Pathology From: beth.villarr...@novartis.com To: histonet@lists.utsouthwestern.edu Date: Thu, 5 Jan 2012 20:02:21 + Subject: [Histonet] saffron vs. safran du gatinais Hello histonet, I have a protocol that calls for safran du gatinais and am experiencing some serious sticker shock. Can I substitute saffron in my solution or am I asking for trouble? Many thanks, Beth Beth Villarreal Scientist I Novartis Institutes for BioMedical Research, Inc. 300 Technology Square Cambridge, MA 02139 USA ___ 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] antibody to CpiV
Is anyone aware of a source for an antibody to CpiV - canine parainfluenza virus. I have searched and come across some papers with IHC staining but I am unable to access the complete paper. Any help would be appreciated. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.comhttp://www.premierlab.com Ship to address: 1567 Skyway Drive, Unit E Longmont, CO 80504 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: saffron vs. safran du gatinais
Hi Beth, Saffron is a great spice. We use it in a old family recipe from Cornwall for a bread roll( saffron nubbies). We also find the price very expensive, but also varies a lot. If you know anyone in the middle East, or India, they could get it a lot cheaper. We have raised it in our garden and get about four 3/4 in long stigmas from each flower, so it is labor intensive to get much, but a little goes a long way. It has to. I don't know of any substitute in staining. Tom Truscott -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Villarreal, Beth Sent: Thursday, January 05, 2012 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] saffron vs. safran du gatinais Hello histonet, I have a protocol that calls for safran du gatinais and am experiencing some serious sticker shock. Can I substitute saffron in my solution or am I asking for trouble? Many thanks, Beth Beth Villarreal Scientist I Novartis Institutes for BioMedical Research, Inc. 300 Technology Square Cambridge, MA 02139 USA ___ 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] Respirators and Routine Histology
I also do not wear any type of respirator. Not at any point of my day. I annually wear a formalin badge to test for exposure rate, but thats it. I gross under a fume hood and I use Slide Bright instead of xylene. It does not have any fumes or noxious odor and is non toxic. My stain line is also contained under a fume hood. No one ever smells or complains about my fumes, unless im changing the processor, they tend to smell the alcohol then. Mask are a required safety supply, and I do believe in some situations a respirator may be needed, but it is ultimately up to the tech. Besides the lab should have adequite ventilation that a respirator should not be needed for the entire shift, maybe during specfic tasks with high fumes. Nicole Tatum, HT ASCP Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. 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] RE: Amazing
0.5% HCL acid ih 70 % ethanol Rena fail On Thu, Jan 5, 2012 at 11:25 AM, Burton, Lynn lynn.bur...@illinois.govwrote: I still use that. It is 70% with 5mL of hydrochloric acid if making 1L. Lynn Burton Lab Assoc I Animal Disease Lab Galesburg, Il 309-344-2451 From: histonet-boun...@lists.utsouthwestern.edu [ histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart [ sdys...@mirnarx.com] Sent: Thursday, January 05, 2012 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Amazing I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Respirators and Routine Histology
I'm in Texas and have been in histology labs since 1998 in 4 different places. None of them did we ever wear a respirator for normal histology work. We did have to wear masks when cutting frozens because of possible TB in lung tissue, but that was it. Most labs should have some kind of negative pressure in them and be exhausting out of the room. I also have always had some kind of hood over the staining station, and usually another hood to gross specimens in. I would say wearing a respirator every day for 8 hours would have made me decide on another field!! How irritating!! Good Luck! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of jsjurc...@comcast.net Sent: Thursday, January 05, 2012 2:31 PM To: Linda Blazek Cc: histonet@lists.utsouthwestern.edu; Amy Self Subject: Re: [Histonet] Respirators and Routine Histology They must be in a liberal part of Michigan. - Original Message - From: Linda Blazek lbla...@digestivespecialists.com To: Lisa Brenner li...@hollandhospital.org, Amy Self as...@georgetownhospitalsystem.org, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Thursday, January 5, 2012 2:30:20 PM Subject: RE: [Histonet] Respirators and Routine Histology Nothing at all??? -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Lisa Brenner Sent: Thursday, January 05, 2012 3:22 PM To: Amy Self; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Respirators and Routine Histology No, not here. We don't wear anything at all. We have excellent air flow, and we wear the badges that test for exposure to xylene and formalin annually which I think is a CAP requirement. Our results have been better than acceptable every year. Lisa Brenner HTL (ASCP) Histology Technical Consultant Holland Hospital phone: (616)394-3184 li...@hollandhospital.org Amy Self as...@georgetownhospitalsystem.org 1/5/2012 3:09 PM Happy New Year to All, I need some help from all of you out there in histoland. How many of you wear respirators during your entire 8 hour work day for routine histology? If you don't wear a respirator do you wear any type of mask or shield at all for routine histology? Also if any of you have any histology safety procedures or information that you would be willing to share with me I would greatly appreciate it. Thanks in advance for all of your help, Amy Amy Self Georgetown Hospital System 843-527-7179 NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: The information contained in this e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information, or Protected Health Information as such term is defined under the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Any unauthorized review, use, disclosure, copying or distribution is prohibited and may be unlawful. If you believe you have received this e-mail in error, please contact the sender by reply e-mail and delete all copies of the original message, including 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 ___ 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: Elastichrome paper, and troubleshooting
How old is the alcoholic Hx you use to prepare the Verhoeffs. I have found that matured 10% ethanoic Hx is more resistant to differentiation than freshly prepared Hx. Might be of use Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy Sent: Friday, 6 January 2012 4:45 AM To: Histonet Subject: [Histonet] Elastichrome paper, and troubleshooting Does anyone have a pdf (or could fax me) the original paper for Elastichrome stain: Richardson, L. Combination Elastic Trichrome Stain, Laboratory Medicine, 6.1, 1975 We have a procedure with this reference, but no original paper to look at. We do it very rarely and no one seems to know how well it worked in the past. I do have another paper in the same vein, A combination verhoeff's elastic and masson's trichrom stain for routine histology, O'Connor, S. Valle, Stain Technology V 57, no. 4, 1982, that is a modification of the Richardson paper, and gives us some ideas, but I would still like to see the original paper if possible. Our problem is that the elastic stain washes out during or after the trichrome. Procedure: Bouins, 56C one hour Wiegerts hematolylin, 3min Verhoffs elastic stain, 15 min 2% ferric chloride differentiation (so far so-good) Gomori's trichrome Blue, 15 min 0.2% glacial acetic acid, 1 min Wash dH20, dehydrate, clear, mount Results: trichrome looks great, no elastin stain. I'm thinking the acetic acid is too long so told them to shorten it to a few dips and see how it looks. But, maybe some has some experience with this and has other suggestions. Thanks for any help. Tim Morken Supervisor, Histology, IPOX UC San Francisco Medical Center Box 1656 1600 Divisidero St, B217 San Francisco, CA 94115 USA 415.514.6042 (office) 415.885.7409 Fax tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet * This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] saffron vs. safran du gatinais
Beth, In the past, people replied to Histonet with the suggestion to buy saffron aka safran du gatinais from a grocery store spice section. Depending on where you are located e.g. a bigger city, try to find a store that sells spices from India may have the freshest saffron. However, you won't suffer the sticker shock of buying it from a chemical supplier. It still tends to be expensive but not like chemical company prices.Once we made the alcoholic saffron solution for Movat's pentachrome, we stored this solution in a container with a dessicant to maintain a water free environment. I was fascinated that Tom actually grew and harvested saffron from the flowers. That is true devotion, but I suspect it was for those delicious sounding nubbies . Good luck Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Villarreal, Beth Sent: Thursday, January 05, 2012 1:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] saffron vs. safran du gatinais Hello histonet, I have a protocol that calls for safran du gatinais and am experiencing some serious sticker shock. Can I substitute saffron in my solution or am I asking for trouble? Many thanks, Beth Beth Villarreal Scientist I Novartis Institutes for BioMedical Research, Inc. 300 Technology Square Cambridge, MA 02139 USA ___ 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: Histonet Digest, Vol 98, Issue 6
Sarah, Your background is caused by cross-activity from your mouse primary antibody on mouse tissue (xenografts). You can try Mouse on Mouse kit (Biocare's has a great Mouse on Mouse HRP Polymer system). I have used Cell Signaling's Rabbit anti-Cleaved Caspase 3 (Cat # 9661) on Xenografts and they work very well. They are very specified and high affinity (appx. ~ 1:10k) with Dako's Envision + system. Your problem will be solve if you just switch your primary ab from mouse to rabbit. Good Luck! Madeleine Huey BS, HTL (ASCP) QIHC Supervisor - Pathology (IPOX Histology) madeleineh...@elcaminohospital.org On Thu, Jan 5, 2012 at 10:00 AM, histonet-requ...@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than Re: Contents of Histonet digest... Today's Topics: 1. RE: decal solution for molecular studies (Clare Thornton) 2. Amazing (Sarah Dysart) 3. RE: Amazing (Bartlett, Jeanine (CDC/OID/NCEZID)) 4. Re: Amazing (Rene J Buesa) 5. RE: Amazing (Bernice Frederick) 6. RE: Amazing (Burton, Lynn) 7. Look at me... (Sarah Dysart) 8. Nail Polish sealant (gayle callis) 9. RE: Look at me... (Elizabeth Chlipala) 10. Elastichrome paper, and troubleshooting (Morken, Timothy) 11. RE: Elastichrome paper, and troubleshooting (Elizabeth Chlipala) -- Message: 1 Date: Thu, 5 Jan 2012 10:16:06 -0500 From: Clare Thornton cthorn...@dahlchase.com Subject: [Histonet] RE: decal solution for molecular studies To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Message-ID: c9d78ffc9d668b4cbea4405f84697504f819b6f...@iris.dahlchase.net Content-Type: text/plain; charset=us-ascii To clarify, the Formical we use is a formic acid/EDTA solution. Clare J. Thornton, HTL(ASCP), QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Clare Thornton Sent: Thursday, January 05, 2012 9:12 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] decal solution for molecular studies Does anyone know of a decal solution that enables molecular studies (Her 2 FISH, EGFR by PCR) to be performed later? We use Formical for our decal solution. Her 2 FISH works about 50% of the time, EGFR almost never. We are looking at having to cut undecalcified bone today for both these studies, and we are not equipped to cut undecalcified bone. Any suggestions? Clare J. Thornton, HTL(ASCP), QIHC Assistant Histology Supervisor Dahl-Chase Diagnostic Services 417 State Street, Suite 540 Bangor, ME 04401 cthorn...@dahlchase.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Message: 2 Date: Thu, 5 Jan 2012 15:59:35 + From: Sarah Dysart sdys...@mirnarx.com Subject: [Histonet] Amazing To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: 8a70a9b2ecdd084dacfe6c59fcf86d509c9...@sn2prd0702mb098.namprd07.prod.outlook.com Content-Type: text/plain; charset=us-ascii I find it amazing sometimes when you don't do something for awhile how quickly your brain throws the information away. That being said...I know back in the day when I was learning histology we used to make our own acid alcohol solution (now where I am had a butt load of Clearifier so I was using that up). I don't want to buy that stuff anymore as making the solution is way cheaper and works just as well. I want to say it was like a 1% acid solution in alcohol?? What was the acid? For some reason my brain says glacial acetic...but time has made me forget. Is the alcohol you mix it in 100% or something lower with a water content to it? Please help my alzheimers =) Happy Thursday!! Sarah Goebel-Dysart, BA, HT(ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 -- Message: 3 Date: Thu, 5 Jan 2012 16:08:36 + From: Bartlett, Jeanine (CDC/OID/NCEZID) j...@cdc.gov Subject: [Histonet] RE: Amazing To: Sarah Dysart sdys...@mirnarx.com, histonet@lists.utsouthwestern.edu
Re: [Histonet] Respirators and Routine Histology
When I work with xylene, it's always in the fume hood while wearing gloves. The same goes for formaldehyde. I never use a face shield for anything, I just do it in the fume hood if I need to use caution with a certain chemical. Emily The whole point of this country is if you want to eat garbage, balloon up to 600 pounds and die of a heart attack at 43, you can! You are free to do so. To me, that’s beautiful. --Ron Swanson ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
