[ aminoreactive dye problem in 8 color panel ]

I would like to share one problem that I realized when I analyzed 
the staining with the aqua-fluorescent vivid dye (excitation: 405 nm and
emission: 525 nm) in combination with an 8 color labeling of PBMC
(pacific blue, fitc, PE, PC5, ECD, PC7 APC and APC-alexa 750).
Occasionally in some stainings, vivid-negative cells in the presence
of this dye  are largely spread over the vivid own channel of 
analysis as well as over some other channels, from violet to PC5, while PC7 
and red laser channels are OK. The vivid dye alone does not seem to be
excited by 488 laser so the fluorescence detected in each channel 
seem to correspond to the antibody used,however, the aspect of cells is
largely spread and makes analysis difficult. Because this dye reacts
also with amines on the cell surface, I wonder whether the
cell-surface staining is the source of these problems. I would 
greatly appreciate any comment on that.

-- 
Françoise Gondois-Rey
IFR 137-CRCM/U891
27 bd Lei Roure 
13009 Marseille
tel 04 91 75 84 21
fax 04 91 26 03 64
f.rey@marseille.inserm.fr