Limit of detection with confocal microscopy compared to flow cytometry

[Klaus Koefoed, Senior Scientist PhD, Antibody Discovery, Symphogen A/S]

Hi:
I was wondering if anyone in this group has done experiments
determining the sensitivity of confocal systems (we use Opera LX)
compared to e.g. flow cytometry. In particular I would like to know
the limit of detection in terms of the minimal cell receptor / antigen
density required for detection of a primary antibody with at secondary
conjugate. I realize of cause that such comparisons are highly
dependent on the reagents used. Just an estimate would be nice.


I have previously worked on the Applied Biosystems 8200 cellular
detection system. The limit of detection of this system has been
reported to be 9000 receptors / cell in a homogenous format (Lee et
al. J Biomol Screen.2008; 13: 210-217).

If anyone has some experience on how to increase the sensitivity
(particular fluorochrome conjugates/polymers, TSA signal
amplification, plate types etc) tt would be high appreciated.
Unfortunately the quantum dot products that are listed on the
Invitrogen webpage are not compatible with the Lasers in our current
Opera LX setup.

Thanks for your time.

Sincerely

Klaus