[Histonet] Need a procedure

[RAY KOELLING]

Tony has great point.  I used to use Histogel (research for cell lines but applicable to cell block).  Switched to Low-melt agarose because of the temp consideration on unfixed cells.  Low-temp is a little misleading.  Melts (in oven at over 60 degrees) but COOL IT and remains liquid at 37degrees (body temp) and some do not gel until 26 degrees.  RNA/DNA-ase free so molecular is fine.  Temp low 37 degrees (when used to mix cells) so no DNA denaturation and proteins fine.  That was 10 years ago. Just checked there are all sorts of "low-melt" agarose out there.

Ray, Fair Director, Eastern Washington Regional Science and Engineering Fair.

> On January 23, 2020 at 2:59 PM "Tony Henwood (SCHN) via Histonet" <histonet at lists.utsouthwestern.edu> wrote:
> 
> 
> Hi all,
> 
> Be careful of using cell block matrix that requires heat to solubilise the matrix (eg agar or other commercial matrixes like Histogel).
> Adding a heated matrix to unfixed, or even formalin fixed, material  can denature some antigens (eg CEA) resulting in a false negative IPX.
> Unfortunately the importance of heat as a pre-analytical factor in immunohistochemistry is often not appreciated.
> 
> 
> 
> 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: John Garratt via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
> Sent: Friday, 24 January 2020 8:21 AM
> To: Muhammad Azam <ajju33 at gmail.com>
> Cc: histonet at lists.utsouthwestern.edu
> Subject: Re: [Histonet] Need a procedure
> 
> http://www.avantec.fr/content/dam/tfs/SDG/APD/APD%20Documents/Product%20Manuals%20&%20Specifications/Histology%20Equipment%20and%20Supplies/Embedding%20Cassettes%20and%20Molds/92957066-Richard-Allan-Scientific-HistoGel-Instructions-for-Use.pdf
> 
> 
> The above link will help.
> 
> 
> 
> www.cpqa.ca
> 
> ‐‐‐‐‐‐‐ Original Message ‐‐‐‐‐‐‐
> On Thursday, January 23, 2020 12:13 PM, Muhammad Azam <ajju33 at gmail.com> wrote:
> 
> > Anybody has validated procedure for histogel
> >
> > Sent from my iPhone
> >
> > > On Jan 23, 2020, at 1:07 PM, John Garratt via Histonet histonet at lists.utsouthwestern.edu wrote:
> > > Hi Terri, I suggest you use Histogel for block preparation. It works exceptionally well, it is good for IHC and does not have the pitfalls of plasma/thrombin.
> > > Plasma/thrombin does work well for cell blocks but you will have to consider an ethical and safe source for your plasma.
> > > The instructions for using Histogel are in the package insert though I have one comment. Be careful how you warm the Histogel and use a heat block. Do NOT use a microwave since there is a tendency to overheat the gel and you will end up with poor quality IHC.
> > > John
> > > www.cpqa.ca
> > > ‐‐‐‐‐‐‐ Original Message ‐‐‐‐‐‐‐
> > >
> > > > On Thursday, January 23, 2020 6:10 AM, Terri Braud via Histonet histonet at lists.utsouthwestern.edu wrote:
> > > > Hi fellow Histonetters - I'm in need of some help, please 
> > > > Background - We currently use agar to capture our scant cell 
> > > > blocks for processing. I am unfamiliar with the Plasma/Thrombin method of cell block preparation and am interested in comparing it to our current method Request - Could you please send me your procedures for this method, specifically where you purchase your plasma and thrombin and what species are used?
> > > > Thanks in advance. Histotechs rock!
> > > > Terri L. Braud, HT(ASCP)
> > > > Anatomic Pathology Supervisor
> > > > Laboratory
> > > > Holy Redeemer Hospital
> > > > 1648 Huntingdon Pike
> > > > Meadowbrook, PA 19046
> > > > ph: 215-938-3689
> > > > fax: 215-938-3874
> > > > Care, Comfort, and Heal
> > > > Histonet mailing list
> > > > Histonet at lists.utsouthwestern.edu
> > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> > >
> > > Histonet mailing list
> > > Histonet at lists.utsouthwestern.edu
> > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> 
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