Dear Paul, Anne and flow users,
Due to the multiple inquiries regarding the feasibility of BD
Pharmingen's Cytometric Bead Array (CBA) products on Coulter's machines, I
would like to take this opportunity to share some technical tips regarding
this issue.
The BD Cytometric Bead Array (CBA) products are compatible with
Coulter XL instruments and the data generated on the XL can be analyzed
using the PC version of the BD CBA software. There are a couple specific
modifications that need to be made to ensure that the software can properly
analyze the list mode data.
It is essential that all data be saved in an FCS 2.0 compliant file
format. The software was designed to work with System 2.0 or higher, and
was not tested on instruments using EXPO Furthermore, the parameters saved
must be in the following order: FSC, SSC, FL1 (FITC), FL2 (PE), and FL3 (or
FL4, bead channel). All parameters should be collected in LOG mode. The
attached word document is designed to help an accomplished XL user with
creating an instrument setup template for the XL and setting up the
instrument as described in the CBA manuals.
If anyone has questions with or information on ways to improve
functionality of CBA products on any flow cytometer we appreciate the input
about our products. Please send your e-mail to me directly at
jfyu@pharmingen.com
(See attached file: Coulter CBA Setup)
Sincerely,
Jifeng Yu, M.D.
Technical Service Scientist
BD Pharmingen & Transduction laboratories.
"PAUL HALLBERG" <Paul.Hallberg@mail.tju.edu> on 08/17/2001 08:01:53 AM
To: cyto-inbox
cc:
Subject: Re: Cytokine bead assay from Pharmingen
Hi Anne,
I am currently working with our BD rep on this same issue. We have
successfully analyzed the results on our PC using their CBA software, but
the samples were acquired on our FACS Calibur. I can't understand why the
software doesn't analyze the data acquired on the Coulter XL. I will share
my solution when resolved.
Regards,
Paul Hallberg
CORE FLow Cytometry Facility
Kimmel Cancer Center
Thomas Jefferson University
-----Original Message-----
From: Anne C Avery [mailto:anne.avery@colostate.edu]
Sent: Wednesday, August 15, 2001 4:27 PM
To: cyto-inbox
Subject: Cytokine bead assay from Pharmingen
Hello,
I was wondering if anyone has used the new cytokine bead assay from
BD/Pharmingen (mouse Th1/Th2 kit) to quantify cytokines. In
particular, all of the support materials address data analysis with BD
instruments and software (not surprisingly). We have a Coulter XL, and
do not have CellQuest. I would be intereested in hearing from anyone
who as anlalyzed the data generated with these reagents as to whether
it can be done using software other than CellQuest and the package
available from Pharmingen (which apparently only works with CellQuest).
Anyone from BD is welcome to weigh in...
Tbanks very much.
Anne
====================================
Anne Avery, VMD, PHD
Department of Pathology
Colorado State Univ
Ft. Collins, CO 80523-1671, USA
voice:(970)491-1170
fax:(970)491-0603
anne.avery@colostate.edu
====================================
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