[Histonet] RE: Rush bx schedule?

[Tony Henwood (SCHN)]

Hi Tim,

Try fixing for 45 min at 50oC. I think it possible that the tissue is not adequately fixed.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior 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: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Morken, Timothy
Sent: Thursday, 5 December 2013 4:31 AM
To: Histonet
Subject: [Histonet] Rush bx schedule?

For tissue, what step most contributes to subsequent swelling of tissue when the block is soaked? I'm thinking the 100% ETOH, xylene clearing and paraffin infiltration all contribute, but at what percentage?

Here is the current schedule, which apparently was meant to mimic a microwave schedule that was used before the MW quit working. The tissue - kidney and liver bx for the most part, up to 10 per day, is coming out soft and swells a bit when soaked. So sectioning is difficult.

We need to have a rapid schedule but need good sections. Obviously we are pushing the limits on this so need a bit of leeway for variation in tissue

So, I'm considering which of these steps is the most critical to lengthen.
This is on a VIP5 but we can also use a peloris for this processing

Formalin 2 min, 50degC (this is not necessary at all, IMHO) 80% ETOH 5 min, 50degC 95%ETOH 5 min, 50degC 100% ETOH 8 min, 50degC Xylene, 9 min 50degC Paraffin, 4 min, Paraffin, 5 min

Thanks for your insight!



Tim Morken
Supervisor, Electron Microscopy and Neuromuscular Special Studies UC San Francisco Medical Center Box 1656
505 Parnassus Ave
San Francisco, CA 94143
USA

415.353.1266  (office)
tim.morken <@t> ucsfmedctr.org<mailto:tim.morken <@t> ucsfmedctr.org>


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