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First some details :
FCM work is done with a Coulter Elite, equipped with a sorter.
Let's define a filamentous alga as 2.5 X 150 micron.
1. When stops a measurement? When I measure time-of-flight I get a sharp
edge at about 40 micron. Is this also th case with with peak or
integrated measurements or will the measurement go on until the particle
has passed the flowcell (i.e. the signal is below the triggervalue)?
Because of the hydrodynamical focussing a filament is supposed to pass
longitudinally, so it should take (in this case) hundred and fifty
microns.
2. Has anyone a (rather simple) solutions to make the measurement longer
and the time-of-flight parameter logarithmic. The range we are dealing
with is about some microns to several hundred microns, so th normal
lineair measurement isn't good enough.
3. What about sorting? A long filamentous alga spans a several droplets!
The formation of droplets will be disturbed but is the sorting still OK?
Is the deflection strong enough? Is every 'droplet' (it will probably be
a chain of 'droplets' with the alga in it) charged?
Thanks in advance,
Hans
Hans Hoogveld
Netherlands Institute of Ecology, Centre for Limnology (NIOO-CL)
Flowcytometry
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