Ralph,

When we use BCIP/NBT, we do our color reaction development in the 37
degree C incubator in the dark.  We change the solution frequently
(every several hours, or when the solution turns pink).  We do not lay
the slides flat for their incubation, instead we use the 5-slide plastic
mailers and have them standing upright.  This keeps precipitate from
settling on to the glass.  Many of the chromogens for Alk. Phos. tend to
form a precipitate, and the manufacturers don't recommend filtering.

Your signal appears to be quite strong.  Perhaps you can also decrease
the time in chromogen?

Best of luck to you,

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64133

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