MTT assay

[Marie]

Hello Everyone,

I am trying to adapt the MTT viability assay for use in 6 well plates
(as opposed to the 96 well plates that the protocols are written for).
 I have a few questions that I hope you can help me with.

1:  After solubilizing, is the formazan released into the medium or is
it maintained in the cells?  (ie:  Can the medium be removed and read
in a cuvette style spectrometer?).

2:  Are there any references to this assay being accurate for 3x10^5
cells or more per well?  If so, could you please point in the right
direction to find this reference.

3:  An old laboratory protocol I found calls for using Hanks BSS for
the incubation step, yet I have found 1 report that using BSS's are
not ideal?  Any thoughts/experience with this?

and finally,

4:  Does anyone know of a protocol where this assay has been modified
for use in other than 96 well plates?

Thank you,

Marie