Bascteria sorting ?

Andy Johnson andy at brc.ubc.ca
Mon Mar 4 16:14:56 EST 2002


Hello

For those of you who know the true finer details about flow and fluid
dynamics ... can somebody tell me the calculation to work out the volume
size of droplets.  The mathamatical approach rather than collecting and
counting the volume.

We are using a 50 µm nozzle at 30psi (BD vantage) and trying to single cell
sort bacteria onto  96 well plate.

The first question is how large are the droplets (volume) ?

Can you determine the dilution factor of your samples (bacteria) ?

We are sorting bacteria and triggering on the fluorescence signal to reduce
background noise.  The questions is : at what rate of sample can we assume
that the instrument is detecting a single cell in a droplet ?

Looking at the numbers it would appear that non-fluorescent bacteria must
be in the same droplets, but I may be assuming something incorrectly.

Any help appreciated and further information available if it will help you
with the solution !

Andy





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