Dear Rob,

Dictyostelium cells seem very sensitive to pressure. While most (not all) mammalian cells would resist in FACS machines to pressure set at 13-20 psi, and even at 70 psi for most recent machines, Dictyostelium cells die even at 13 psi.  They survive well at 9 psi, with nozzle diameter set at 100 microns.  We are using FACS-sorting successfully under these conditions, in particular for cell cloning and for selecting fluorescent-most cells.

With regards,

Pierre
____________
Pierre Golstein
Centre d'Immunologie de Marseille-Luminy
INSERM/CNRS/Univ.Medit.
Case 906, Campus de Luminy, Avenue de Luminy,  13288 Marseille cedex 9, France
tel : 33 (0)4 91 26 94 68   Fax : 33 (0)4 91 26 94 30
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http://www.ciml.univ-mrs.fr/

De : Robert Cooper <[log in to unmask]<mailto:[log in to unmask]>>
Répondre à : DICTY <[log in to unmask]<mailto:[log in to unmask]>>
Date : Mon, 29 Aug 2011 18:17:58 -0400
À : <[log in to unmask]<mailto:[log in to unmask]>>
Objet : [DICTY - 630] live cell FACS

I was wondering if anyone has achieved good results for live cell sorting of Dicty cells using FACS.  I can sort cells fine, but it seems that more than 99% of the cells die in the process.  I gather the cells in KK2 and sort them through the FACS machine into another tube of KK2, but less than 1% of the events that were sorted into that tube end up growing into colonies on a bacterial plate.  Does anyone have any techniques or tips for increasing the cells' survival rate through the FACS machine?

Thanks,
~Rob Cooper
Cox lab