We are trying to improve our assay for counting cells in 96 well plates
using a fluorescent microplate reader. We are very lazy and skint (don't
like buying kits) , and would like to add the dye to a well without the need
to centrifuge and remove unbound dye.
The characteristics of such a dye would be the following:
1. Fluorescence enhancement on binding to nucleic acid
2. High affinity for nucleic acid
3. DNA specificity.
4. Economic
In the molecular probes catalogue we are told that Cyquant^TM green kit is
the best dye for this purpose, and can detect down to 50 cells under optimal
conditions, however we would like to find an alternative which is not a kit.
We have tried PI + 1% Triton 100 which goes down to 2000 cells (no wash).
Background fluorescence was very high for H33342 and DAPI (no wash). We were
wondering whether we could should try one of the many new cyanine dyes.
Which nucleic acid stains (other than Cyquant^TM green) would be best for
the job?
?Thiazole orange
?TOTO / YOYO / BOBO/ HOHO^tm
Many thanks for your patents.
Robert Nordon
Post Doctoral Fellow
Graduate School of Biomedical Engineering
University of New South Wales
Sydney, 2052
Tel. no. 61-2-9385-1148
Fax 61-2-9663-2108