Sorting bionanoparticles.

Hello flow'ers.

I am new to the world of flow cytometry and have a client who would like me
to sort bionanoparticles.  I have a BD FACSVantage-DiVa DiVa with Turbo
Sort.  The bionanoparticles range in size from 50-500nm and are lipid based
so will be stained with Nile red.

I would like to know if I can sort the particles that are <200nm if they are
stained with Nile red?	Would it then be possible to sort the ~200-500nm
particles and save the conflicts to get two groups, i.e. ~50-200nm and
~200nm-500nm?  The smallest nozzle I have is the 50µ.	This nozzle has not
been utilised at our institute before, so could anyone give me an indication
of the frequency and pressure to use the 50µ nozzle at?

There is also a possibility of staining the bionanoparticles with Nile red
and another fluorophore if this would aid the sorting process.

One last question:  What filter or bandpass would be best to measure Nile
red emission?  I have found conflicting data regarding the emission spectra
of Nile red bound to lipid.  Practical Flow Cytometry states that the
fluorescence of the lipid-bound dye is about 550nm and another source
(Invitrogen fluorescence spectra viewer) has the excitation max of
lipid-bound dye at 550nm and the emission at 636nm?

I would appreciate any advice/information.

Thanks

Kylie Price

Staff Scientist ­ Flow Cytometry
The Malaghan Institute of Medical Research
The Central Services Building
Victoria University Campus
Entrance 7
Kelburn Parade
Wellington
New Zealand

Ph:	       64-4-4996914, ext 850
Fax:	       64-4-4996915
E-mail:       kprice@malaghan.org.nz
Web:	      www.malaghan.org.nz





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