Re: viability of cells after sorting

From: Daniel Vaulot (vaulot@sb-roscoff.fr)
Date: Mon Jan 27 1997 - 11:36:45 EST


The following references are relevant to phytoplankton cell sorting:

Rivkin RB, Phinney DA, Yentsch CM (1986) Effects of flow cytometric analysis
and cell sorting on photosynthetic carbon uptake by phytoplankton in
cultures and from natural populations. Appl Environ Microbiol 52:935-938; 
Li WKW (1994) Primary productivity of prochlorophytes, cyanobacteria, and
eucaryotic ultraphytoplankton: measurements from flow cytometric sorting.
Limnol Oceanogr 39:169-175; 
Sensen CW, Heinmann K, Melkonian M (1993) The production of clonal and
axenic cultures of micoralgae using fluorescence activated cell sorting. Eur
J Phycol 28:93-97;
Lipschultz F (1995) Nitrogen-specific uptake rates of marine phytoplankton
isolated from natural populations of particles by flow cytometry. Mar Ecol
Prog Ser 123:245-258

In our lab we have been using flow cytometry for almost 10 years to sort
phytoplankton either directly from natural communities or from cultures and
we obtained viable cells in most cases.  Our problem is rather to get pure
cultures after sorting because when we sorted from mixtures of algae we
often failed to recover pure cultures from only one type.

The following recommendations apply.
- Sterilize your instrument before sorting
- If you deal with marine algae, use 0.2 um filtered sea water as sheath fluid
- Rince very thoroughly so that no trace of bleach (if bleach is used for
routine cleaning) is left.
- If you are using an instrument with a big laser, lower the laser power as
much as you can (below 100 mW).


Daniel Vaulot
______________________________________
Daniel VAULOT
Station Biologique CNRS - INSU - UPMC
BP 74 
29682 Roscoff Cx FRANCE

Ph: 33 2 98 29 23 34 Fax: 33 2 98 29 23 24

e-mail: vaulot@sb-roscoff.fr
W3: http://www.sb-roscoff.fr (PhytoPlankton Group)



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