Dear Chlamy community
I'm trying to grow and mate Chlamy in 96-well plates. I'd be curious to
hear from other people who have experience in this area.
My personnal experience with flat bottom plates is that the cells tend
to form a ring at the bottom, and I don't think it's good if you want to
reach decent cell densities or see your cells survive for a decent
period of time. Has anyone compared round-bottom or V-shaped plates ?
Any feelings about plastic ? Polystyrene, vs. polyethylene, vs. special
plastics (e.g. Nunc Microwell low-binding) ? I'd be tempted to go for a
low-binding plastic, because Chlamy likes to swim around, but do the
cheaper plates work ?
Is agitation possible at all in these small wells ? How do you control
evaporation, especially of the outermost wells ?
What about large-volume wells ? I tried 1.3 ml and 2 ml Deepwell plates,
but the cell density remained low so I never gained much in terms of
yield. The surface/volume ratio becomes terrible, and the opaque plastic
reduces light access. The cells tends to fall to the bottom. Has anyone
had more success ?
And mating ? There are quite a few labs out there who do matings in
96-well format, but I'm not sure what type of plate they use, and how
they place the cells in gametogenesis conditions. Does diluting from TAP
into water or N-free medium works ? Has anyone succeeded in storing the
zygotes in the wells ? I myself have used cat litter, but its handling
becomes quite messy in a microtiter plate format. Do glass beads work ?
Thanks for your answers, and other thoughts on this not-so-trivial subject
Olivier Vallon
--
Olivier VALLON
CNRS, UMR 7141
CNRS/Université Pierre et Marie Curie
Institut de Biologie Physico-Chimique
13 rue Pierre et Marie Curie
75005 Paris
FRANCE
ovallon from ibpc.fr
tel: (33) (0)1 5841 5058
fax: (33) (0)1 5841 5022
http://www.ibpc.fr/UMR7141/
