RE: [Histonet] Antibody Validation
Funny, I'd think the vendors would want you to do tons of your own validation, as it uses up antibody faster, ergo they get to sell you more antibody! :) In a feisty mood today. Claire From: histonet-boun...@lists.utsouthwestern.edu on behalf of Rene J Buesa Sent: Tue 6/15/2010 9:49 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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] NY licensing
I'm looking into requirements for getting licensed as a histotech in NY and I am a little unsure of whether my educational experience will be considered a substantial equivalent to their requirements. They require education from a Histology program or something equivalent. I graduated from a 4 year university (UC Davis) with a B.S. in Animal Science (includes all science courses--biology, chem, organic chemistry, physics, anatomy etc) but I never specifically took a class on histology and its techniques used in the lab. Do you still think they will consider me eligible or would they require me to complete a histology program? I have yet to find the chance to call them since they close at 4:45 ET and I am on the west coast. Thanks in advance, Jennifer Campbell ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Fume hood
Brandi- Having gone through and even designed lab renovations, my advice is: Have a different hood for each function if possible. Renovations cannot see into the future to know what technologies you might need later. An extra hood now can very easily seem like too few later. Also, many functions need to be separated, a fact that administrators will never understand. You might need vents over your processors, a vented area for special stains, a hood for grossing and a hood for stainers/ coverslippers. Cytology may need several hoods also. If genetic determination requirements become standard, will this be in your lab? Emergency power for each hood. Also processors and other necessary equipment (stainers, etc.). Desktop height for many histology applications, especially grossing, embedding and sectioning. Otherwise, the ergonomics are terrible. I would suggest backdraft enclosed hoods for grossing, vented covered areas for processors, stainers/coverslippers. If using xylenes, enclose as much as possible and use backdraft. Be sure to keep in mind that with backdraft hoods, there may need to be face opening restrictions to achieve the majic number of 100 cfm you need for safety. If you have anyother questions, please get back with me. Joe Saby, BA HT From: Walter Benton To: histonet@lists.utsouthwestern.edu Sent: Tue, June 15, 2010 12:49:25 PM Subject: [Histonet] Fume hood Brandi, I have been through several renovations and highly recommend that you get a hood that exhaust outside. Depending upon what applications you plan to carry out under the hood, filters may not do the trick. It is also important to have someone from your facilities or engineering department conduct an air exchange analysis to make sure that you have proper airflow and exchanges within the room, since the two operations are being combined. I would also recommend that you have your area monitored for air quality while performing the various tasks that you perform now in you current configuration to see if you are OSHA compliant or whatever regulatory body you wish to follow. Downdraft ventilation is great for Xylene, because Xylene vapor is heavier than air, thus the reason those systems work well. Feel free to reach out if you have any other questions. Walter Benton, HT(ASCP)QIHC Histology Supervisor University of Maryland Medical Center Anatomic Pathology 22 S. Greene St Room NBW65 Baltimore MD 21201 (Direct) 410-328-0930 (Lab) 410-328-5524 (Fax) 410-328-5508 >>> On 6/15/2010 at 12:31 PM, wrote: Message: 13 Date: Tue, 15 Jun 2010 09:51:04 -0400 From: Brandi Higgins Subject: [Histonet] fume hood To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello, Our hospital is doing some renovation and we need to look into new fume hoods for our new location. Currently we have one fume hood over our grossing area, and one fume hood in our coverslipping area (two different rooms). The hospital wants to put our grossing room and histo/cyto rooms together. I am still going to need two separate hoods. Does anyone have any experience/knowledge/input about fume hoods? I'm trying to look into the ductless ones, although I imagine changing the filters will end up being more expensive over time (I have no idea what would be involved in running a duct/vent). Also I have seen a benchtop downdraft type that sucks the air down, and does not have a top. It is advertised as being good for xylene. Does anyone use this in their coverslipping area? Any input would be greatly appreciated. I'm pretty clueless on the whole issue. I want to make sure that what I get will be safe for me and my coworker as we will be spending most of our day in this room. Any input is appreciated! Thank You! Brandi Higgins, BS, HT(ASCP) This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. ___ 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] liquidating lab equipment
Hello Cathy, I can be interessed on these 2 eqs. Shandon paraffin embeddding center with heated forceps VIP 2000 tissue processor Do you have any pictures ? Regards Ricky > Date: Tue, 15 Jun 2010 13:38:12 -0500 > From: camay...@gmail.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] liquidating lab equipment > > I have a few pieces of lab equipment left over from when I closed and > liquidated the lab. I have the following: > > 1) Shandon paraffin embeddding center with heated forceps > 2) VIP 2000 tissue processor > 3) 6 foot fume hood (1/2 of base cabinet is flammable storage) > 4) 2 Well Diamond wire saws (model 4240) > 5) Exakt grinder > > all of the equipment was being used right up to the last week we closed the > lab > > -- > Cathy Mayton > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Hotmail is redefining busy with tools for the New Busy. Get more from your inbox. http://www.windowslive.com/campaign/thenewbusy?ocid=PID28326::T:WLMTAGL:ON:WL:en-US:WM_HMP:042010_2___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] alcohol monitoring
Hi Cristi, We have been monitoring alcohol fumes for many years. Our hospital safety officer orders badges and we wear them on our lab coats for 15 minutes and another one for 8 hours. Only one tech needs to wear them in the lab, not everybody. We usually do it on a day where we will handle more alcohol... Like cleaning the processors and recycling. We also wear the badges for xylene and formalin fumes. Again, one for 15 minutes and another for 8 hours. Once we are done, we put them back in their designated bags and call the safety officer to pick them up. He sends them off to wherever he orders them and they send back the report. Very easy... If you want, I could put you in touch with our safety officer so you can get further information. Good luck, Karen Karen L. Bauer HT/HTL (ASCP) Histology Section Chief Department of Pathology - Luther Hospital Luther Midelfort - Mayo Health System 715-838-3205 bauer.ka...@mayo.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cristi stephenson Sent: Tuesday, June 15, 2010 1:32 PM To: Histo Net Subject: [Histonet] alcohol monitoring Hello, I was recently approached by my management about monitoring the alcohol in the lab? I monitor for xylene and formalin, but I have never heard of doing so for alcohol. Does anyone else do this at their lab? Is there potentially a different type of testing to check for this? Any guidance is much appreciated. Thanks, Cristi ___ 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] alcohol monitoring
What type of "alcohol" are you being asked to test and why? OSHA does establish limits for Isopropyl Alcohol (PEL 400 ppm) because it is an irritant. If you are doing tissue processing w/out xylene, then you are probably using isoporpyl alcohol and should test personnel maintaining the processor. I think you will need additional information before you can set a plan. William DeSalvo, B.S., HTL(ASCP) > Date: Tue, 15 Jun 2010 11:32:23 -0700 > From: cls71...@sbcglobal.net > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] alcohol monitoring > > Hello, > I was recently approached by my management about monitoring the alcohol in > the lab? I monitor for xylene and formalin, but I have never heard of doing > so for alcohol. Does anyone else do this at their lab? Is there potentially > a different type of testing to check for this? Any guidance is much > appreciated. > Thanks, > Cristi > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ The New Busy think 9 to 5 is a cute idea. Combine multiple calendars with Hotmail. http://www.windowslive.com/campaign/thenewbusy?tile=multicalendar&ocid=PID28326::T:WLMTAGL:ON:WL:en-US:WM_HMP:042010_5___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] alcohol monitoring
Never done it, never requested, really unnecessary. Perhaps the "requester" was trying to "push his/her pencil" harder than others in creating a new layer of bureaucratic work to just be "more busy"! René J. --- On Tue, 6/15/10, Cristi stephenson wrote: From: Cristi stephenson Subject: [Histonet] alcohol monitoring To: "Histo Net" Date: Tuesday, June 15, 2010, 2:32 PM Hello, I was recently approached by my management about monitoring the alcohol in the lab? I monitor for xylene and formalin, but I have never heard of doing so for alcohol. Does anyone else do this at their lab? Is there potentially a different type of testing to check for this? Any guidance is much appreciated. Thanks, Cristi ___ 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] Part-Time Job Opening in Portland, OR
Part-Time Laboratory Technician Part-time laboratory technician needed for private histology laboratory. Job duties include accessioning and handling medical specimens. Applicant should be familiar with human anatomy, have attention to detail, and be self motivated. HTL/HT (or equivalent) encouraged to apply. Bachelors degree in biology, chemistry or other basic science with a minimum GPA 3.2 required. Salary DOE Email kate...@medsurgpath.com or fax resumes to (503) 670-9754 Katelin Lester, HTL(ASCP) MedSurg Pathology Associates, Inc. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] alcohol monitoring
Hello, I was recently approached by my management about monitoring the alcohol in the lab? I monitor for xylene and formalin, but I have never heard of doing so for alcohol. Does anyone else do this at their lab? Is there potentially a different type of testing to check for this? Any guidance is much appreciated. Thanks, Cristi ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] liquidating lab equipment
I have a few pieces of lab equipment left over from when I closed and liquidated the lab. I have the following: 1) Shandon paraffin embeddding center with heated forceps 2) VIP 2000 tissue processor 3) 6 foot fume hood (1/2 of base cabinet is flammable storage) 4) 2 Well Diamond wire saws (model 4240) 5) Exakt grinder all of the equipment was being used right up to the last week we closed the lab -- Cathy Mayton ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Antibody Validation - long response
Well said Elizabeth Jesus A Ellin HT/PA ASCP Department of Pathology/Histology Yuma Regional Medical Center 2400 South Ave A Yuma, AZ 85364 - 7170 ( Office: (928) 336-1743 (Fax: (928) 336-7319 *Email: jel...@yumaregional.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Tuesday, June 15, 2010 9:01 AM To: Rene J Buesa; histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: RE: [Histonet] Antibody Validation - long response Bottom line it's not the vendors responsibility to validate their equipment or antibodies in your lab. Some vendors may help you do this, but ultimately the lab needs to validate the equipment and IHC in their lab. The vendors normally calibrate the equipment prior to shipment and once they set the instrument up in your lab, they should be able to provide you with the documentation that states that they calibrated the instrument. Your instruments need to be calibrated prior to being validated. As far as your scanner goes some vendors can provide validation, but it's at a cost and that cost is not cheap depending upon what you actually want validated. If you are using the scanner and associated algorithms for analysis then you need to validate that separately. There are several steps required to validate a scanner - 1. you validate the scanner 2. if you are using a database to store your images then that also may need to be validated and 3. if you are using algorithms that provide you with data then those algorithms need to be validated. For example prior to running a validation protocol on a tissue processor its needs to be calibrated for temperature. All of your major equipment needs to be on a calibration schedule. We calibrate all of our instruments once a year and validation is completed only once unless we change the instrument location or how we use the instrument. Pipettors are calibrated every 6 months. All instruments are validated it may just be a one pager for the basic lab equipment but instruments like the tissue processor, slide staining, IHC stainer and scanner require written protocols some of these are 80 pages in length and go into great detail. The same goes for your antibodies. Antibodies are validated initially with 25 tissue samples (10 strongly positive tissues, 10 moderate to weakly positive tissues and 5 tissues that have no reactivity) This type of validation is required for routine antibodies, prognostic markers such as Her-2, ER and PR require additional tissue samples. New lots require 3 tissue samples one strongly positive on moderate to weakly positive and one negative. If you change the antibody source or detection system or retrieval it needs to be validated again - This information comes from the paper Standarization of Immunohistochemistry from CAP its available on line - I have a copy if you need it. There are also new guidelines for ER/PR and a new article on validation of ER/PR in the June issue of Archives of pathology from CAP. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, June 15, 2010 8:50 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request f
[Histonet] Fume hood
Brandi, I have been through several renovations and highly recommend that you get a hood that exhaust outside. Depending upon what applications you plan to carry out under the hood, filters may not do the trick. It is also important to have someone from your facilities or engineering department conduct an air exchange analysis to make sure that you have proper airflow and exchanges within the room, since the two operations are being combined. I would also recommend that you have your area monitored for air quality while performing the various tasks that you perform now in you current configuration to see if you are OSHA compliant or whatever regulatory body you wish to follow. Downdraft ventilation is great for Xylene, because Xylene vapor is heavier than air, thus the reason those systems work well. Feel free to reach out if you have any other questions. Walter Benton, HT(ASCP)QIHC Histology Supervisor University of Maryland Medical Center Anatomic Pathology 22 S. Greene St Room NBW65 Baltimore MD 21201 (Direct) 410-328-0930 (Lab) 410-328-5524 (Fax) 410-328-5508 >>> On 6/15/2010 at 12:31 PM, wrote: Message: 13 Date: Tue, 15 Jun 2010 09:51:04 -0400 From: Brandi Higgins Subject: [Histonet] fume hood To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello, Our hospital is doing some renovation and we need to look into new fume hoods for our new location. Currently we have one fume hood over our grossing area, and one fume hood in our coverslipping area (two different rooms). The hospital wants to put our grossing room and histo/cyto rooms together. I am still going to need two separate hoods. Does anyone have any experience/knowledge/input about fume hoods? I'm trying to look into the ductless ones, although I imagine changing the filters will end up being more expensive over time (I have no idea what would be involved in running a duct/vent). Also I have seen a benchtop downdraft type that sucks the air down, and does not have a top. It is advertised as being good for xylene. Does anyone use this in their coverslipping area? Any input would be greatly appreciated. I'm pretty clueless on the whole issue. I want to make sure that what I get will be safe for me and my coworker as we will be spending most of our day in this room. Any input is appreciated! Thank You! Brandi Higgins, BS, HT(ASCP) This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] fume hood
Hi Brandi, Three years ago, our lab was totally renovated. Clinical and anatomic pathology have become one big room. The only rooms that are separate are the grossing room (who sports a nifty Mopec 600 grossing w/vented hood system) and the TB lab. There has been some adjusting for the employees during this time. The med techs could not tolerate our xylene under any circumstances. Just changing the solutions in the processor/stainers was a whole new experience for them, much less any minor spill. In fact, having a minor spill ended up closing the lab for service, but that is another story. Switching over to xlyene substitute has worked out well after making adjustments. It sounds like just you and cytology are combined, so I hope that cytology has considered a hood for their non-gyn prep as you just never know about contaminates. Our cytology area has a great biohazard floor model and is also vented. In histology, we have a back draft built right into the back of the countertops that helps a lot with our manual special stains, and our automated coverslipper (era 1994) has a cover over it and is tied into the same venting system as the grossing station. In retrospect, the only changes I wish I would have thought of, was to make sure that the entire venting system has a dedicated electrical hook-up. Because when the electricity goes down (and it will due to weather or a frisky squirrel), people cannot work in the fumes. It's amazing how quickly it gets bad. At first you don't realize it and then everyone starts getting xylene/formalin side affects. Not pretty. That's all I can think of and sorry it was long winded! Try and get the best for you and your people. Filters are expensive and in my opinion, don't give as good air exchange and a vented system. Best regards, Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor MSH Competency Coordinator Hurley Medical Center One Hurley Plaza Flint, MI 48503 email: lpave...@hurleymc.com ph: 810-257-9948 Lab: 810-257-9138 fax: 810-762-7082 >>> Rene J Buesa 6/15/2010 10:53 AM >>> Over the counter fumes hood with filters are very effcicient and cheaper than running the vent ducts, especially in a large building. They also have the advantage that they can be moved to another place if necessary. Those without a cover are not that efficient. René J. --- On Tue, 6/15/10, Brandi Higgins wrote: From: Brandi Higgins Subject: [Histonet] fume hood To: histonet@lists.utsouthwestern.edu Date: Tuesday, June 15, 2010, 9:51 AM Hello, Our hospital is doing some renovation and we need to look into new fume hoods for our new location. Currently we have one fume hood over our grossing area, and one fume hood in our coverslipping area (two different rooms). The hospital wants to put our grossing room and histo/cyto rooms together. I am still going to need two separate hoods. Does anyone have any experience/knowledge/input about fume hoods? I'm trying to look into the ductless ones, although I imagine changing the filters will end up being more expensive over time (I have no idea what would be involved in running a duct/vent). Also I have seen a benchtop downdraft type that sucks the air down, and does not have a top. It is advertised as being good for xylene. Does anyone use this in their coverslipping area? Any input would be greatly appreciated. I'm pretty clueless on the whole issue. I want to make sure that what I get will be safe for me and my coworker as we will be spending most of our day in this room. Any input is appreciated! Thank You! Brandi Higgins, BS, HT(ASCP) ___ 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] Antibody Validation - long response
Bottom line it's not the vendors responsibility to validate their equipment or antibodies in your lab. Some vendors may help you do this, but ultimately the lab needs to validate the equipment and IHC in their lab. The vendors normally calibrate the equipment prior to shipment and once they set the instrument up in your lab, they should be able to provide you with the documentation that states that they calibrated the instrument. Your instruments need to be calibrated prior to being validated. As far as your scanner goes some vendors can provide validation, but it's at a cost and that cost is not cheap depending upon what you actually want validated. If you are using the scanner and associated algorithms for analysis then you need to validate that separately. There are several steps required to validate a scanner - 1. you validate the scanner 2. if you are using a database to store your images then that also may need to be validated and 3. if you are using algorithms that provide you with data then those algorithms need to be validated. For example prior to running a validation protocol on a tissue processor its needs to be calibrated for temperature. All of your major equipment needs to be on a calibration schedule. We calibrate all of our instruments once a year and validation is completed only once unless we change the instrument location or how we use the instrument. Pipettors are calibrated every 6 months. All instruments are validated it may just be a one pager for the basic lab equipment but instruments like the tissue processor, slide staining, IHC stainer and scanner require written protocols some of these are 80 pages in length and go into great detail. The same goes for your antibodies. Antibodies are validated initially with 25 tissue samples (10 strongly positive tissues, 10 moderate to weakly positive tissues and 5 tissues that have no reactivity) This type of validation is required for routine antibodies, prognostic markers such as Her-2, ER and PR require additional tissue samples. New lots require 3 tissue samples one strongly positive on moderate to weakly positive and one negative. If you change the antibody source or detection system or retrieval it needs to be validated again - This information comes from the paper Standarization of Immunohistochemistry from CAP its available on line - I have a copy if you need it. There are also new guidelines for ER/PR and a new article on validation of ER/PR in the June issue of Archives of pathology from CAP. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, June 15, 2010 8:50 AM To: histo...@pathology.swmed.edu; teri.hall...@midmichigan.org Subject: Re: [Histonet] Antibody Validation Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the r
RE: [Histonet] Antibody Validation
There are other issue that you have to take into consideration with both instruments, With digital image analysis you also have to look at the questions that CAP just instituted for this. This includes the machine being run by someone that can do high complexity testing. This means that you have to have a tech do this machine, not a TA or Lab aid. There are also issue with consistent calibration of the Image analysis machines, continued education of the techs, and validating not only anitbodies but continued validation of the image algorythms.. Do not listen to vendors, rely on your instinct and validate. Jesus A Ellin HT/PA ASCP Department of Pathology/Histology Yuma Regional Medical Center 2400 South Ave A Yuma, AZ 85364 - 7170 ( Office: (928) 336-1743 (Fax: (928) 336-7319 *Email: jel...@yumaregional.org __ This message is confidential, intended only for the named recipient(s) and may contain information that is privileged or exempt from disclosure under applicable law. If you are not the intended recipient(s), you are notified that the dissemination, distribution, or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at either the e-mail, fax, address, or telephone number listed above and delete this e-mail from your computer. Thank You. __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody Validation
I agree with Rene. All lot to lot and new antibodies need to be checked for consistency. This is part of your validation process. This even holds true for pre-dilutes which are already tested by the manufacturer for optimum results. Beatrice Sullivan, HT(A.S.C.P.) HTL , AAS, CLSP(N.C.A.) AP Supervisor Shore Memorial Hospital 609-653-3590 Rene J Buesa To Sent by: histo...@pathology.swmed.edu, histonet-bounces@ teri.hall...@midmichigan.org lists.utsouthwest cc ern.edu Subject Re: [Histonet] Antibody Validation 06/15/2010 10:49 AM Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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] Uneven fluorescent staining
Thank you so much for your answer Anatoli! - I tried the citrate buffer boiling system and I obtained the same unreliable results with NeuN. Some animals showed a great NeuN staining and others did not. I really thought at that point that my perfusion was the problem and I switched from making my own PFA4% to buying a 16%PFA from Electron Microscopy Sciences, which I think solved the problem. Now I'm obtaining a good NeuN staining but only in some regions of the tissue. - I am using the Rat monoclonal from Abcam (1:250) - I'll consider the new method using Edu labeling for new experiment and will try the ProLong Gold antifade, thank you very much for the advice. Alexia Nunez University of Maryland, College Park > Subject: RE: [Histonet] Uneven fluorescent staining > Date: Tue, 15 Jun 2010 10:26:32 -0400 > From: agleiber...@cbiolabs.com > To: alexia_f...@hotmail.com > > Alexia, > 1. HCl treatment could be sometimes problematic - many epitopes are quite > sensitive to it, so your NeuN antibody will work not in optimal conditions. > You can replace acid treatment with citrate buffer boiling - the same > treatment that is used for paraffin sections. It will denature DNA as > effectively as HCl treatment. > 2. You did not mention what antibody you are using for BrdU - the best are > rat monoclonal from Abcam (much better than any mouse monoclonal or sheep or > goat polyclonal I have tested) - and you can easy combine them with mouse > monoclonal against NeuN (I believe, only mouse anti-NeuN exist). > 3. For future experiments I recommend switch from BrdU labeling to EdU > labeling (Click-iT® EdU Alexa Fluor® 488 HCS Assay from Invitrogen) - this > technique is more sensitive for studying DNA synthesis than BrdU, staining is > faster (30min), does not involve neither denaturation step nor antibody and > can be easily combined with any other marker staining. To do antigen staining > after EdU visualization, that is simple chemical reaction, you need wash > thoroughly your slides and perform your antibody staining. Additional benefit > is - much better structure of nuclei because denaturation step sometimes > affects them. In addition, because of high sensitivity you can decrease > concentration of nucleotide substitute (EdU) 5-10 times in comparison with > BrdU that can be crucial for long-term experiments because all these > nucleotide analogues (especially BrdU) can affect cell fate in long run. > 4. Finally, I recommend to replace Vectashield with ProLong Gold antifade > (Invitrogen) with or without DAPI. It is better for fluorochrome preservation > and it will harden overnight - you don't need any nail polish > > Anatoli Gleiberman, PhD > Director of Histopathology > Cleveland Biolabs, Inc > 73 High Street > Buffalo, NY 14203 > phone:716-849-6810 ext.354 > fax:716-849-6817 > e-mail: agleiber...@cbiolabs.com > > -Original Message- > From: histonet-boun...@lists.utsouthwestern.edu > [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Alexia > Francisca Núñez Parra > Sent: Monday, June 14, 2010 6:10 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Uneven fluorescent staining > > > Hello everyone!! > The Histonet Archives have been always very helpful to me, so I decided to > become a member and ask a personal question to the experts. > > I am doing NeuN and BrdU double staining in the olfactory bulb, following the > protocol described below. I have always had some trouble with the NeuN > staining, obtaining sometimes good or no staining at all. I tried to trouble > shoot with the intracardial perfusion and even bought a new NeuN antibody > (Chemicon). I am having now, however, another problem. I observed an uneven > NeuN and BrdU staining. There are some parts of the section that get a really > good and bright staining and others that do not get any staining at all (even > though they should). > > Can anyone give a hint on what to review or change? > > Thank you so much!!! > > Alexia > > 1. Intracardial perfusion with cold PBS and cold 4%PFA. > 2. The brains are postfixed for 4 hrs in PFA 4% at 4C and immersed in sucrose > 30% ON. After that, the brains are embedded in tissue tek and store at -80. > 3. Sections of 20um are obtained using a cryostat and collected using > positive charged slides. > > > > > > > > > > > > > > > 4. > Allow the slides to reach the RT > > > 5. > Warm slides at 55C x 10' > > 6. > Outline slides with Immedge pen > > 7. > Hydrate: PBS 2 x 5', final quick wash with ddH20 > > 8. > Soak slides in 2N HCl for 1 hr at 37C > > 9. > Neutralize washing the slides with 0.1M Na tretaborate buffer > pH8.5, 3 x 10' > > 10. > Wash with PBS 2 x 10' > > 11. > Wash with PBS-T 1 x 10' > > > > 12. > Block at RT with 10% of Donkey serum in PBST x 2hrs (60uL on each section) > > 13. > First antibody: incubate at 4C with first antibody diluited in > PBST
RE: [Histonet] fume hood
There are two units we use in our labs and I suggest looking into the Mopec BF600 Downdraft Workcell. This works great for H&E/FS stain lines and other areas w/ open containers of chemicals. I also suggest considering the Labconco Protector XVS Ventilation Station for counter top hood with easy access, the unit is connected to outside air extraction. There are many other options and you should do a search on the internet for more options. William DeSalvo, B.S., HTL(ASCP) Production Manager, Sonora Quest laboratories Chair, NSH QCC > Date: Tue, 15 Jun 2010 09:51:04 -0400 > From: brandihigg...@gmail.com > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] fume hood > > Hello, > > Our hospital is doing some renovation and we need to look into new fume > hoods for our new location. Currently we have one fume hood over our > grossing area, and one fume hood in our coverslipping area (two different > rooms). The hospital wants to put our grossing room and histo/cyto rooms > together. I am still going to need two separate hoods. Does anyone have > any experience/knowledge/input about fume hoods? I'm trying to look into > the ductless ones, although I imagine changing the filters will end up being > more expensive over time (I have no idea what would be involved in running a > duct/vent). Also I have seen a benchtop downdraft type that sucks the air > down, and does not have a top. It is advertised as being good for xylene. > Does anyone use this in their coverslipping area? Any input would be > greatly appreciated. I'm pretty clueless on the whole issue. I want to > make sure that what I get will be safe for me and my coworker as we will be > spending most of our day in this room. Any input is appreciated! Thank > You! > > Brandi Higgins, BS, HT(ASCP) > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ The New Busy is not the old busy. Search, chat and e-mail from your inbox. http://www.windowslive.com/campaign/thenewbusy?ocid=PID28326::T:WLMTAGL:ON:WL:en-US:WM_HMP:042010_3___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Antibody Validation
Teri: You are right about the validations you propose although I am not surprised that your vendor does not think it is necessary. They are in the business of selling and you are in the business of assuring the high quality of your work to obtaining the most accurate work for patients' sake. There is where the difference resides. Ignore your vendor and keep validating your protocols. René J. --- On Tue, 6/15/10, teri.hall...@midmichigan.org wrote: From: teri.hall...@midmichigan.org Subject: [Histonet] Antibody Validation To: histo...@pathology.swmed.edu Date: Tuesday, June 15, 2010, 7:55 AM I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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] fume hood
Over the counter fumes hood with filters are very effcicient and cheaper than running the vent ducts, especially in a large building. They also have the advantage that they can be moved to another place if necessary. Those without a cover are not that efficient. René J. --- On Tue, 6/15/10, Brandi Higgins wrote: From: Brandi Higgins Subject: [Histonet] fume hood To: histonet@lists.utsouthwestern.edu Date: Tuesday, June 15, 2010, 9:51 AM Hello, Our hospital is doing some renovation and we need to look into new fume hoods for our new location. Currently we have one fume hood over our grossing area, and one fume hood in our coverslipping area (two different rooms). The hospital wants to put our grossing room and histo/cyto rooms together. I am still going to need two separate hoods. Does anyone have any experience/knowledge/input about fume hoods? I'm trying to look into the ductless ones, although I imagine changing the filters will end up being more expensive over time (I have no idea what would be involved in running a duct/vent). Also I have seen a benchtop downdraft type that sucks the air down, and does not have a top. It is advertised as being good for xylene. Does anyone use this in their coverslipping area? Any input would be greatly appreciated. I'm pretty clueless on the whole issue. I want to make sure that what I get will be safe for me and my coworker as we will be spending most of our day in this room. Any input is appreciated! Thank You! Brandi Higgins, BS, HT(ASCP) ___ 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] fume hood
Hello, Our hospital is doing some renovation and we need to look into new fume hoods for our new location. Currently we have one fume hood over our grossing area, and one fume hood in our coverslipping area (two different rooms). The hospital wants to put our grossing room and histo/cyto rooms together. I am still going to need two separate hoods. Does anyone have any experience/knowledge/input about fume hoods? I'm trying to look into the ductless ones, although I imagine changing the filters will end up being more expensive over time (I have no idea what would be involved in running a duct/vent). Also I have seen a benchtop downdraft type that sucks the air down, and does not have a top. It is advertised as being good for xylene. Does anyone use this in their coverslipping area? Any input would be greatly appreciated. I'm pretty clueless on the whole issue. I want to make sure that what I get will be safe for me and my coworker as we will be spending most of our day in this room. Any input is appreciated! Thank You! Brandi Higgins, BS, HT(ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE Antibody validation
Teri New lots of Antibodies recieved by a lab should be validated by that laboratory. Thus proving the new lots produce similar results sa the old under the same set of circumstances. Processing, Retrevial methods, thickness of sections, techniques etc used in your laboratory contribute to the outcome. Having the Ab validated by the vendor does not verify that it works in your laboratory with your procedures. Rena Fail - Original Message From: "teri.hall...@midmichigan.org" To: histo...@pathology.swmed.edu Sent: Tue, June 15, 2010 7:55:28 AM Subject: [Histonet] Antibody Validation I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ 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 Validation
I am being questioned by our vendor as to why we need to validate our automated immunostainer and image analysis instrument. They would like documentation pertaining to the requirement of validation and the number of specimens utilized for validation. I am requesting that each antibody be validated on the instrument against a previously validated instrument. Additionally, I am requesting that each new lot of antibody be validated upon receipt against previously ran specimens. This would also apply to the image analysis antibodies. (Her2 has been validated by FISH.) The vendor has apparently polled users in the area and this is not a standard protocol, therefore the request for documentation. I think it is pretty clearly stated by CAP in the Quality Management In Anatomic Pathology. Any other suggestions? Teresa Hallada BS, MT/CT (ASCP) Pathology Lead MidMichigan Health - Gratiot teri.hall...@midmichigan.org 989.463.1101 ext 3423 ___ Please note that this email message and any attachments may contain privileged and confidential information that is protected against use or disclosure under federal and state law. The information is intended only for the personal and confidential use of the intended recipient. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this information in error and that any review, dissemination, distribution, copying or action taken in reliance on the contents of this communication is strictly prohibited. If you have received this email in error, please advise by immediate reply. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
