Re: [Histonet] Dirty H after break
Hi Allan, With the quality of the eosin dyes today, filtering should not be required. The same for xylene and ethanols. You will need to monitor the usage and solution levels (evaporation being the issue here). There are no hard and fast rules but a suggestion is one slide per ml of solution. Therefore a bath of 400ml of haematoxylin should stain 400 slides consistently before a noticeable deterioration in staining is noted. The same with the other solutions. 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: Allan Wang [mailto:alla...@gmail.com] Sent: Saturday, 7 January 2017 4:01 PM To: Tony Henwood (SCHN) Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dirty H after break Yep, filtering the hematoxlyn fixed it. Thanks for the tip. Is it worthwhile to filter the eosin, alcohols, and xylene substitutes as well? Allan On Jan 4, 2017 1:04 AM, "Tony Henwood (SCHN)" <tony.henw...@health.nsw.gov.au<mailto:tony.henw...@health.nsw.gov.au>> wrote: Hi Allan, The flakes are alum precipitates. I would suggest filtering the haematoxylin before use. 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 -Original Message- From: Allan Wang via Histonet [mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>] Sent: Wednesday, 4 January 2017 4:45 PM To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: [Histonet] Dirty H after break Hello all, Does someone know what would cause dirty flakes of light purple that showed across the whole slide? http://i.imgur.com/1nIXwAy.jpg Reagents are relatively fresh but the H stainer hadn't been used for a week over the holiday break. Is it surface film on the hematoxylin? I didn't notice anything in the container. We are using Mayer's, followed by differentiation with HCl acid alcohol, with 2-3 minute long wash steps in between. Thanks, Allan ___ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto: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. 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] Dirty H after break
Yep, filtering the hematoxlyn fixed it. Thanks for the tip. Is it worthwhile to filter the eosin, alcohols, and xylene substitutes as well? Allan On Jan 4, 2017 1:04 AM, "Tony Henwood (SCHN)" < tony.henw...@health.nsw.gov.au> wrote: > Hi Allan, > > The flakes are alum precipitates. > > I would suggest filtering the haematoxylin before use. > > 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 > > -Original Message- > From: Allan Wang via Histonet [mailto:histonet@lists.utsouthwestern.edu] > Sent: Wednesday, 4 January 2017 4:45 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Dirty H after break > > Hello all, > > Does someone know what would cause dirty flakes of light purple that > showed across the whole slide? > > http://i.imgur.com/1nIXwAy.jpg > > Reagents are relatively fresh but the H stainer hadn't been used for a > week over the holiday break. > Is it surface film on the hematoxylin? I didn't notice anything in the > container. We are using Mayer's, followed by differentiation with HCl acid > alcohol, with 2-3 minute long wash steps in between. > > Thanks, > Allan > ___ > 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] Dirty H after break
Hi Allan, The flakes are alum precipitates. I would suggest filtering the haematoxylin before use. 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 -Original Message- From: Allan Wang via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Wednesday, 4 January 2017 4:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dirty H after break Hello all, Does someone know what would cause dirty flakes of light purple that showed across the whole slide? http://i.imgur.com/1nIXwAy.jpg Reagents are relatively fresh but the H stainer hadn't been used for a week over the holiday break. Is it surface film on the hematoxylin? I didn't notice anything in the container. We are using Mayer's, followed by differentiation with HCl acid alcohol, with 2-3 minute long wash steps in between. Thanks, Allan ___ 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
[Histonet] Dirty H after break
Hello all, Does someone know what would cause dirty flakes of light purple that showed across the whole slide? http://i.imgur.com/1nIXwAy.jpg Reagents are relatively fresh but the H stainer hadn't been used for a week over the holiday break. Is it surface film on the hematoxylin? I didn't notice anything in the container. We are using Mayer's, followed by differentiation with HCl acid alcohol, with 2-3 minute long wash steps in between. Thanks, Allan ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
