Decrease in MFI using HTS on LSRII

Morschauser, Andrew andrew.morschauser at roche.com
Tue Jun 5 11:27:33 EDT 2007


Dear Group,

 

I am posting the following question for a colleague:

 

Here is my question for the Flow cytometry group.

 

 

I have been trying to develop a plate based assay to measure a
cell-surface antigen on CHO cells using the LSRII-HTS system.

 

I perform the staining in 12X75mm polypropylene tubes.	At the end of
the staining procedure, I transfer to polystyrene tubes and the mean
fluorescence of intensity is determined on the LSRII-HTS system.  I then
take that same sample, transfer to polypropylene or polystyrene U-bottom
96-well plates and re-analyze the samples using the HTS system.

 

The MFI of the samples analyzed in the tubes is > than	the MFI of the
samples analyzed in the plates?  It is the same sample.

Shouldn't the MFI's be relatively the same?

 

 I would like this to be a plate-based assay because there are quite a
number of samples to be evaluated but I don't feel comfortable with the
plate-based results.

 

Any hints or suggestions would be greatly appreciated.  

 

 

Sincerely,

 

Andy Morschauser

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