[Cytometry] Pleural effusion sample prep for sorting

Donnenberg, Vera donnenbergvs at upmc.edu
Fri May 6 09:34:59 EDT 2011


Dear Kathy-
We have done many PEs, including a mesothelioma, and published extensively in this matter.  Here are some protocols for your review.  Basically, each PE has to be heparinized, washed (save the cell-free fluid for chemokine analysis) and collagenase/DNAse/Dispase digested (tumors are in microclumps), followed by a 2mM EDTA-0.5% Human Serum Albumin PBS buffer suspension. We use 100uM nozzle, keep the sort at 4C.  If you are doing Hoechst transport, do not chealate calcium.
Let me know if you need more info.  Good luck.
Vera
VSD
Assistant (412) 623-7780/7781
Office (412) 623-7784
Laboratory (412) 623-3266
FAX (412) 623-7778


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Kathy Heel
Sent: Wednesday, May 04, 2011 11:07 PM
To: Purdue Flow MailList (cytometry at lists.purdue.edu)
Subject: [Cytometry] Pleural effusion sample prep for sorting

Hello All,

We've had a request to sort pleural effusions from mesothelioma patients and was wondering if anyone has any experience with these samples and suggestions for sample prep? I've been told the effusions can be very rich in protein and coagulating agents and can be very sticky.

Many thanks
Kathy

Kathy Heel PhD
Assistant Professor
Centre for Microscopy, Characterisation and Analysis (M510)
The University of Western Australia
35 Stirling Highway
Crawley WA 6009
Australia

Ph 61 8 9346 4525
Fax 61 8 9346 3469
Mob 0416 085 960
www.cmca.uwa.edu.au<http://www.cmca.uwa.edu.au>

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