Re: [Histonet] "cooked" biopsy
Thank you for the insights into recycling. Our surveys are based on labs achieving a quality of staining and processing that ensures accurate clinical interpretation and labs are generally doing an excellent job regardless of the reagents used which is a tribute to their QC. It would be great to see more standardization in histotechnology with the increased reliance on anatomic pathology to guide therapy, but I suspect that is a pipe dream. Here is an article that may be of interest https://link.springer.com/article/10.1007/s00428-020-02960-z John On Fri, Jul 9, 2021 at 7:18 PM, Jamie Watson wrote: > Hello everyone, > > Tony is correct that generally recycled xylene is purer than what you buy. I > have been using recycled xylene since the early 80s from 4 different types of > recyclers. > > The best practice is to have a small amount of new xylene constantly coming > into the recycling stream. We always use purchased xylene on our H stainer > to achieve this. Also about every 1-1.5 years replace all the xylene in the > recycling stream. > > Alcohol recycling is too labor intensive and can be difficult to get the the > percentage correct. > > I have never used recycled formalin. > > Jamie > > On July 9, 2021 6:13:13 PM "Tony Henwood (SCHN) via Histonet" > wrote: > >> Hi John, >> >> There is anecdotal evidence that recycled xylene is of a better quality than >> the original purchased product. We have found it to be so. No processing nor >> staining problems unless there has been a recycling issue (as shown by the >> CBG xylene purity test). >> >> I suppose we have to access each reagent that is recycled and determine the >> risk of using it. For example, unless care is taken, recycling alcohol can >> be an issue (eg xylene contamination). We do not recycle alcohol for this >> reason. >> >> "When we are continuously challenged by pre-analytic variables in >> histotechnology why do labs continue to use recycled reagents? " >> >> From your survey, has re-cycling xylene been found to be a major, or minor >> issue? >> >> We definitely need evidence that re-cycling is a risk. >> >> Regards >> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) >> Principal Scientist, the Children’s Hospital at Westmead >> Adjunct Fellow, School of Medicine, University of Western Sydney >> Tel: 612 9845 3306 >> Fax: 612 9845 3318 >> Pathology Department >> the children's hospital at westmead >> Cnr Hawkesbury Road and Hainsworth Street, Westmead >> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA >> >> From: John Garratt >> Sent: Saturday, 10 July 2021 05:31 >> To: Paula; Tony Henwood (SCHN); 'Erick Rodriguez' >> Cc: histonet@lists.utsouthwestern.edu >> Subject: Re: [Histonet] "cooked" biopsy >> >> Interesting discussion. >> At CPQA we recently started a H EQA program that includes fixation and >> processing in the feedback to participants and in a recent mini-survey I >> found that a third of labs use recycled product somewhere in the >> pre-analytic phase. >> When we are continuously challenged by pre-analytic variables in >> histotechnology why do labs continue to use recycled reagents? >> >> John >> >> On Thu, Jul 8, 2021 at 5:59 AM, Paula via Histonet >> mailto:histonet@lists.utsouthwestern.edu>> >> wrote: >> Hi Tony, >> Thank you for the procedure. I do send out the recycled xylene to have its >> analysis done by an outside company every month, and I'll put into place >> this testing procedure that you outlined below in place ase well. >> Paula >> >> -Original Message- >> From: Tony Henwood (SCHN) [mailto:tony.henw...@health.nsw.gov.au] >> Sent: Wednesday, July 07, 2021 1:25 PM >> To: 'Erick Rodriguez'; Paula >> Cc: histonet@lists.utsouthwestern.edu >> Subject: Re: [Histonet] "cooked" biopsy >> >> Hi Paula, >> >> We can check the purity of the xylene quite easily: >> >> Xylene Purity Test Procedure >> >> Note: The recommended and most accurate method of determining the purity of >> the recycled xylene is by doing a Gas Chromatography analysis. The following >> method can be used to obtain an acceptable confidence level in the purity of >> the recycled xylene (CBG Biotech). >> >> Testing Procedure >> >> 1. To a clean, dry 100 ml mixing cylinder graduate, add sufficient recovered >> xylene so that the bottom of the meniscus is aligned with the top edge of &g
Re: [Histonet] "cooked" biopsy
Hi John, There is anecdotal evidence that recycled xylene is of a better quality than the original purchased product. We have found it to be so. No processing nor staining problems unless there has been a recycling issue (as shown by the CBG xylene purity test). I suppose we have to access each reagent that is recycled and determine the risk of using it. For example, unless care is taken, recycling alcohol can be an issue (eg xylene contamination). We do not recycle alcohol for this reason. "When we are continuously challenged by pre-analytic variables in histotechnology why do labs continue to use recycled reagents? " >From your survey, has re-cycling xylene been found to be a major, or minor >issue? We definitely need evidence that re-cycling is a risk. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: John Garratt Sent: Saturday, 10 July 2021 05:31 To: Paula; Tony Henwood (SCHN); 'Erick Rodriguez' Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Interesting discussion. At CPQA we recently started a H EQA program that includes fixation and processing in the feedback to participants and in a recent mini-survey I found that a third of labs use recycled product somewhere in the pre-analytic phase. When we are continuously challenged by pre-analytic variables in histotechnology why do labs continue to use recycled reagents? John On Thu, Jul 8, 2021 at 5:59 AM, Paula via Histonet mailto:histonet@lists.utsouthwestern.edu>> wrote: Hi Tony, Thank you for the procedure. I do send out the recycled xylene to have its analysis done by an outside company every month, and I'll put into place this testing procedure that you outlined below in place ase well. Paula -Original Message- From: Tony Henwood (SCHN) [mailto:tony.henw...@health.nsw.gov.au] Sent: Wednesday, July 07, 2021 1:25 PM To: 'Erick Rodriguez'; Paula Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Hi Paula, We can check the purity of the xylene quite easily: Xylene Purity Test Procedure Note: The recommended and most accurate method of determining the purity of the recycled xylene is by doing a Gas Chromatography analysis. The following method can be used to obtain an acceptable confidence level in the purity of the recycled xylene (CBG Biotech). Testing Procedure 1. To a clean, dry 100 ml mixing cylinder graduate, add sufficient recovered xylene so that the bottom of the meniscus is aligned with the top edge of the 85 ml mark on the graduate. 2. Add water to the graduate until the bottom of the meniscus aligns with the top edge of the 100 ml mark on the graduate. At this point, 15 ml of water will have been added to 85 ml of recovered xylene. 3. Stopper the graduate and invert the mixture. Allow the mixture to settle, making sure that all of the water settles to the bottom of the graduate. No water should remain clinging to the sides of the graduate above the xylene/water separation point. This separation point should be near the 15 ml level of the graduate. (Note: xylene floats on top of the water). 4. Carefully inspect and record the point of separation between the water and xylene using the bottom of the meniscus as the separation point. 5. Subtract 15 ml from the quantity of water indicated in step 5. The remainder plus an additional 0.1 correction factor equals the percentage of recovered xylene impurities. EXAMPLE: Xylene/Water separation point is indicated to be 15.5 ml. (15.5 - 15) + 0.1 = 0.6% impurities. Therefore, the recovered xylene is 99.4% pure. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: Paula via Histonet Sent: Thursday, 8 July 2021 04:53 To: 'Erick Rodriguez' Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Thank you, everyone... I looked at my reagents and saw the color pink in the xylene, which tells me that there is water or too much water in it so I changed it. We recycle xylene, so I need to get the recycler looked at now. Thanks again, Paula -Original Message- From: Erick Rodriguez [mailto:rodriguez.er...@icloud.com] Sent: Wednesday, July 07, 202
Re: [Histonet] "cooked" biopsy
Interesting discussion. At CPQA we recently started a H EQA program that includes fixation and processing in the feedback to participants and in a recent mini-survey I found that a third of labs use recycled product somewhere in the pre-analytic phase. When we are continuously challenged by pre-analytic variables in histotechnology why do labs continue to use recycled reagents? John On Thu, Jul 8, 2021 at 5:59 AM, Paula via Histonet wrote: > Hi Tony, > Thank you for the procedure. I do send out the recycled xylene to have its > analysis done by an outside company every month, and I'll put into place this > testing procedure that you outlined below in place ase well. > Paula > > -Original Message- > From: Tony Henwood (SCHN) [mailto:tony.henw...@health.nsw.gov.au] > Sent: Wednesday, July 07, 2021 1:25 PM > To: 'Erick Rodriguez'; Paula > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "cooked" biopsy > > Hi Paula, > > We can check the purity of the xylene quite easily: > > Xylene Purity Test Procedure > > Note: The recommended and most accurate method of determining the purity of > the recycled xylene is by doing a Gas Chromatography analysis. The following > method can be used to obtain an acceptable confidence level in the purity of > the recycled xylene (CBG Biotech). > > Testing Procedure > > 1. To a clean, dry 100 ml mixing cylinder graduate, add sufficient recovered > xylene so that the bottom of the meniscus is aligned with the top edge of the > 85 ml mark on the graduate. > > 2. Add water to the graduate until the bottom of the meniscus aligns with the > top edge of the 100 ml mark on the graduate. At this point, 15 ml of water > will have been added to 85 ml of recovered xylene. > > 3. Stopper the graduate and invert the mixture. Allow the mixture to settle, > making sure that all of the water settles to the bottom of the graduate. No > water should remain clinging to the sides of the graduate above the > xylene/water separation point. This separation point should be near the 15 ml > level of the graduate. (Note: xylene floats on top of the water). > > 4. Carefully inspect and record the point of separation between the water and > xylene using the bottom of the meniscus as the separation point. > > 5. Subtract 15 ml from the quantity of water indicated in step 5. The > remainder plus an additional 0.1 correction factor equals the percentage of > recovered xylene impurities. > > EXAMPLE: > > Xylene/Water separation point is indicated to be 15.5 ml. > (15.5 - 15) + 0.1 = 0.6% impurities. > Therefore, the recovered xylene is 99.4% pure. > > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) > Principal Scientist, the Children’s Hospital at Westmead > Adjunct Fellow, School of Medicine, University of Western Sydney > Tel: 612 9845 3306 > Fax: 612 9845 3318 > Pathology Department > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > ____ > From: Paula via Histonet > Sent: Thursday, 8 July 2021 04:53 > To: 'Erick Rodriguez' > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "cooked" biopsy > > Thank you, everyone... > I looked at my reagents and saw the color pink in the xylene, which tells me > that there is water or too much water in it so I changed it. > We recycle xylene, so I need to get the recycler looked at now. > Thanks again, > Paula > > -Original Message- > From: Erick Rodriguez [mailto:rodriguez.er...@icloud.com] > Sent: Wednesday, July 07, 2021 11:24 AM > To: Paula > Subject: Re: [Histonet] "cooked" biopsy > > Did you change the processor reagents before running your tissues? Cooked > tissue usually means the tissue wasn’t dehydrated properly and the leftover > water boiled and fried your tissue. I would double check the alcohols. > >> On Jul 7, 2021, at 11:00 AM, Paula via Histonet >> wrote: >> >> Hello, good day, >> >> >> >> Our pathologist is complaining about the tissues today that they are >> "cooked, burnt, crushed, shrunken" those are the adjectives she is using. >> >> >> >> Can you tell me the cause? Usually, the work comes out beautiful but today >> they are not. Nothing has changed on our processing times. >> >> >> >> What should I investigate? >> >> >> >> Thank you in advance, >> >> Paula >> >> Bio-Path Medica Group >> >> >> >> ___
Re: [Histonet] "cooked" biopsy
Hi Tony, Thank you for the procedure. I do send out the recycled xylene to have its analysis done by an outside company every month, and I'll put into place this testing procedure that you outlined below in place ase well. Paula -Original Message- From: Tony Henwood (SCHN) [mailto:tony.henw...@health.nsw.gov.au] Sent: Wednesday, July 07, 2021 1:25 PM To: 'Erick Rodriguez'; Paula Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Hi Paula, We can check the purity of the xylene quite easily: Xylene Purity Test Procedure Note: The recommended and most accurate method of determining the purity of the recycled xylene is by doing a Gas Chromatography analysis. The following method can be used to obtain an acceptable confidence level in the purity of the recycled xylene (CBG Biotech). Testing Procedure 1. To a clean, dry 100 ml mixing cylinder graduate, add sufficient recovered xylene so that the bottom of the meniscus is aligned with the top edge of the 85 ml mark on the graduate. 2. Add water to the graduate until the bottom of the meniscus aligns with the top edge of the 100 ml mark on the graduate. At this point, 15 ml of water will have been added to 85 ml of recovered xylene. 3. Stopper the graduate and invert the mixture. Allow the mixture to settle, making sure that all of the water settles to the bottom of the graduate. No water should remain clinging to the sides of the graduate above the xylene/water separation point. This separation point should be near the 15 ml level of the graduate. (Note: xylene floats on top of the water). 4. Carefully inspect and record the point of separation between the water and xylene using the bottom of the meniscus as the separation point. 5. Subtract 15 ml from the quantity of water indicated in step 5. The remainder plus an additional 0.1 correction factor equals the percentage of recovered xylene impurities. EXAMPLE: Xylene/Water separation point is indicated to be 15.5 ml. (15.5 - 15) + 0.1 = 0.6% impurities. Therefore, the recovered xylene is 99.4% pure. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: Paula via Histonet Sent: Thursday, 8 July 2021 04:53 To: 'Erick Rodriguez' Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Thank you, everyone... I looked at my reagents and saw the color pink in the xylene, which tells me that there is water or too much water in it so I changed it. We recycle xylene, so I need to get the recycler looked at now. Thanks again, Paula -Original Message- From: Erick Rodriguez [mailto:rodriguez.er...@icloud.com] Sent: Wednesday, July 07, 2021 11:24 AM To: Paula Subject: Re: [Histonet] "cooked" biopsy Did you change the processor reagents before running your tissues? Cooked tissue usually means the tissue wasn’t dehydrated properly and the leftover water boiled and fried your tissue. I would double check the alcohols. > On Jul 7, 2021, at 11:00 AM, Paula via Histonet > wrote: > > Hello, good day, > > > > Our pathologist is complaining about the tissues today that they are > "cooked, burnt, crushed, shrunken" those are the adjectives she is using. > > > > Can you tell me the cause? Usually, the work comes out beautiful but today > they are not. Nothing has changed on our processing times. > > > > What should I investigate? > > > > Thank you in advance, > > Paula > > Bio-Path Medica Group > > > > ___ > 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 This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] "cooked" biopsy
Hi Paula, We can check the purity of the xylene quite easily: Xylene Purity Test Procedure Note: The recommended and most accurate method of determining the purity of the recycled xylene is by doing a Gas Chromatography analysis. The following method can be used to obtain an acceptable confidence level in the purity of the recycled xylene (CBG Biotech). Testing Procedure 1. To a clean, dry 100 ml mixing cylinder graduate, add sufficient recovered xylene so that the bottom of the meniscus is aligned with the top edge of the 85 ml mark on the graduate. 2. Add water to the graduate until the bottom of the meniscus aligns with the top edge of the 100 ml mark on the graduate. At this point, 15 ml of water will have been added to 85 ml of recovered xylene. 3. Stopper the graduate and invert the mixture. Allow the mixture to settle, making sure that all of the water settles to the bottom of the graduate. No water should remain clinging to the sides of the graduate above the xylene/water separation point. This separation point should be near the 15 ml level of the graduate. (Note: xylene floats on top of the water). 4. Carefully inspect and record the point of separation between the water and xylene using the bottom of the meniscus as the separation point. 5. Subtract 15 ml from the quantity of water indicated in step 5. The remainder plus an additional 0.1 correction factor equals the percentage of recovered xylene impurities. EXAMPLE: Xylene/Water separation point is indicated to be 15.5 ml. (15.5 - 15) + 0.1 = 0.6% impurities. Therefore, the recovered xylene is 99.4% pure. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA From: Paula via Histonet Sent: Thursday, 8 July 2021 04:53 To: 'Erick Rodriguez' Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "cooked" biopsy Thank you, everyone... I looked at my reagents and saw the color pink in the xylene, which tells me that there is water or too much water in it so I changed it. We recycle xylene, so I need to get the recycler looked at now. Thanks again, Paula -Original Message- From: Erick Rodriguez [mailto:rodriguez.er...@icloud.com] Sent: Wednesday, July 07, 2021 11:24 AM To: Paula Subject: Re: [Histonet] "cooked" biopsy Did you change the processor reagents before running your tissues? Cooked tissue usually means the tissue wasn’t dehydrated properly and the leftover water boiled and fried your tissue. I would double check the alcohols. > On Jul 7, 2021, at 11:00 AM, Paula via Histonet > wrote: > > Hello, good day, > > > > Our pathologist is complaining about the tissues today that they are > "cooked, burnt, crushed, shrunken" those are the adjectives she is using. > > > > Can you tell me the cause? Usually, the work comes out beautiful but today > they are not. Nothing has changed on our processing times. > > > > What should I investigate? > > > > Thank you in advance, > > Paula > > Bio-Path Medica Group > > > > ___ > 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 This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of NSW Health or any of its entities. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] "cooked" biopsy
Thank you, everyone... I looked at my reagents and saw the color pink in the xylene, which tells me that there is water or too much water in it so I changed it. We recycle xylene, so I need to get the recycler looked at now. Thanks again, Paula -Original Message- From: Erick Rodriguez [mailto:rodriguez.er...@icloud.com] Sent: Wednesday, July 07, 2021 11:24 AM To: Paula Subject: Re: [Histonet] "cooked" biopsy Did you change the processor reagents before running your tissues? Cooked tissue usually means the tissue wasn’t dehydrated properly and the leftover water boiled and fried your tissue. I would double check the alcohols. > On Jul 7, 2021, at 11:00 AM, Paula via Histonet > wrote: > > Hello, good day, > > > > Our pathologist is complaining about the tissues today that they are > "cooked, burnt, crushed, shrunken" those are the adjectives she is using. > > > > Can you tell me the cause? Usually, the work comes out beautiful but today > they are not. Nothing has changed on our processing times. > > > > What should I investigate? > > > > Thank you in advance, > > Paula > > Bio-Path Medica Group > > > > ___ > 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
