Re: BD CBA on a Coulter XL

From: tward@PharMingen.com
Date: Mon Nov 06 2000 - 12:56:44 EST


Hi  Wayne and others who may be interested.

Yes the BD CBA can be read on a Coulter XL.  Coversion of Coulter CBA files via
FACSConvert will put the files to FCS 2.0 in the right format for the MacIntosh.
The CBA software MS Excel Add-in on the mac platform will automatically detect
the instrument source (ie Coulter XL in this case) and process the files as it
would normally for a BD generated file.

It is critical that the MS Excel has the solver add-in installed in addition to
the CBA add in.  Unfortunately the universality of MS Excel across platforms is
not universal.  We run into problems running this on PC MS Excel installs.  We
have not been successful to date in tracking down the inconsistencies between
mac/pc performance of the add - in. This means that the analysis of the FCS
files must be done on a mac version of excel.

Re: BD CBA on an XL:


The Coulter XL flow cytometer is able to use either FL3 with minor modification
or the optional FL4 unmodified to run the BD Cytometric Bead Array. For FL3
approaches, a standard 660/40 nm band pass from Omega Optical can be installed
on the FL3 detector and obtain excellent resolution and low level of noise
compared to the standard bandpass of 620/30 . If the FL4 detector option is
installed on the Coulter XL, then the CBA particles can be run directly with no
modification to the optical components required.



Our information shows that a set up voltage within 900 to 1,000 volts be used
with the FL-3 detector fitted with a 660/40 bandpass filter to optimally
minimize background.  The use of the 660/40 bandpass in the FL3 position is
critical in resolving the 7 bead mix, and provides for excellent resolution and
control of noise levels while maintaining a decent voltage (900 to 1,000 volts
out of 2,000).



For the 4th detector option, testing of the set up bead mix using voltages of
800, 850 and 900 on the FL4 detector indicate that a voltage setting of around
850 on the FL4 detector and using the supplied bandpass filter, the target value
(500) as indicated can be easily attained.


Hope this helps

Regards

Tony Ward
BD Biosciences



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