From owner-celegans@net.bio.net Sun Aug 17 23:00:00 1997 Path: biosci!agate!newsfeed.kornet.nm.kr!news.maxwell.syr.edu!newsfeed.sovam.com!sovam!demos!Gamma.RU!srcc!newsserv From: "Vladimir V. Bakaev" Newsgroups: bionet.celegans Subject: Re: C. elegans in microtitre wells, aging Date: 18 Aug 1997 07:08:25 +0400 Organization: Private Person Lines: 42 Distribution: world Message-ID: Reply-To: vlad@bakaev.nsk.ru NNTP-Posting-Host: news.gamma.ru Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Summary: Growth in microtitre wells Keywords: C. elegans X-Return-Path: bakaev!bakaev.nsk.ru!vlad@sinor.ru 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. .