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C. elegans in microtitre wells, aging

Vladimir V. Bakaev vlad at bakaev.nsk.ru
Sun Aug 17 22:08:25 EST 1997


In his letter on 08 Aug 1997 Ralf Baumeister wrote:
>Hi,
>I saw your worm breeder's gazette articles and wanted to ask you some
>question concerning growing worms in small volumes of microtiter plates:
>We have been trying to grow worms in microtiter plates because of space
>problems in the incubators. However, our worms died mostly before they
>had progeny.
>There are several observations we made:
>1. The plastic of the microtiter plates seem to make a difference.
>2. Aeration seems to be a critical parameter.
>Since you are an expert in this field, I would like to ask you for your
>experience. In particular:
>1. How do your aerate your plates (rotation, shaking at which speed)?
>2. Is there a particular material/brand of microtiter plate you recommend?
>3. What is the volume of liquid you use, and which medium and bacteria
>   do you propose?
>4. How do you prevent evaporation of liquid from the wells during long
>   incubations (10 days and more)?
>Thank you in advance for your answer.

>Ralf Baumeister

     Dear Dr.  Ralf Baumeister,  I am sorry for a long silence,  but I
was in another place. Now I am trying to answer:
     I am working only with microtitre wells which are called here  as
immunological plates (23x12cm),  they have 12x6 wells,  every well has
volume 2,5ml. I do not know the name of this plastic, but nematodes in
my experiments have maximal lifespan comparable with this parameter in
experiments of another authors.
     I use  0,75ml  of  liquid  S medium in each well.  This medium is
described in the book THE NEMATODE CAENORHABDITIS ELEGANS (William  B.
Wood, ed.),  1988,  p.590. Earlier I used bacteria E. coli OP 50 only,
but I found that it is possible to use in my experiments any other  E.
coli. I add bacteria E. coli to liquid S medium.
     To prevent evaporation I place any plate on another, etc.,and up-
per plate is empty. As for aeration, _I every day transfer the animals
in the following well with fresh medium_.
     Thank you for your letter and I'll be glad to answer on your pos-
sible questions.
     Yours truly, Vladimir.
     RUSSIA, 630107, Novosibirsk, box 45, Bakaev V.V.




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