...nystatin works well at concentrations recommended for mammalian cells.
We never had problems with yeasts through contaminated media from Formedium.
All the best, Ralph
Am 02.07.2013 um 13:32 schrieb ranjani dhakshinamoorthy:
> Hello all,
> I have a question on this regard. Even after preparing the Dicty cultures at aseptic conditions if we end up getting yeast contamination for unknown reasons is it a good idea to use the antifungal agent Nystatin, which is suggested for cell culture work?
>
> Thanks,
> Ranjani
>
> --
> Ranjani Dhakshinamoorthy,
> C/o Dr. Baskar,
> Dept. of Biotechnology,
> Indian Institute of Technology Madras,
> Chennai, INDIA
>
>
> -----------------------------------------------------------------------------------------------------------------------------------------------
>
> On Tue, Jul 2, 2013 at 2:57 PM, Professor Paul Fisher <[log in to unmask]> wrote:
> Dear Charlotte
>
> Yeast should not survive autoclaving (not even as spores), but your filters might be contaminated. I would not bother with filtering.
> The other possibility is that the autoclaving conditions are not right. Standard would be 121C for 20 min but if you are using large volumes, the heating in the central areas of the container will be slower and so may not reach the desired temperature early enough.
>
> Also watch out that any post-autoclaving manipulations, additions, inoculations might introduce contaminants.
>
> Cheers,
> Paul.
>
> Prof. Paul R. Fisher,
> Chair in Microbiology,
> Deputy Executive Dean,
> Faculty of Science, Technology & Engineering,
> La Trobe University,
> VIC 3086,
> AUSTRALIA.
>
> Tel. + 61 3 9479 2229
> Fax. + 61 3 9479 1222
> Email. [log in to unmask]
> Web. http://fishfam.homeip.net:100/mcbg
>
> La Trobe University - ranked top in Victoria for student satisfaction
> (Sweeney Uni Student Report, 2009)
>
>
> On 2 July 2013 16:31, Pierre Cosson <[log in to unmask]> wrote:
> Dear Charlotte,
> What happened to us once: we were using so-called sterile bacteriological plates for Dictyo cultures, and started getting low level infections from time to time, often with yeast. After checking everything, the answer was that the sterile bacteriological plates are not truly sterile. They are produced in a sterile environment, but some lots will contain a few spores. There are two solutions: 1-quick and expensive: use tissue-culture plastic; beware, this will change the behavior of your cells, since Dictyo adheres much better to tissue-culture plastic. 2-cheap: order a lot of bacteriological plates that have been sterilized by irradiation. That costs very little, and solves completely the problem. The only problem is that you need to order a full batch of plates (I think minimum 10-20 boxes).
> If you are already using tissue-culture plastic, then maybe consider checking the filter of your culture hood. Anyway, autoclaved HL5, filtered or not, is sterile, so this is not the cause of your problems.
> Best regards
> Pierre
>
>
> Pierre Cosson
> Centre Médical Universitaire
> Dpt of Cell Physiology and Metabolism
> 1 rue Michel Servet
> CH 1211 Geneva 4, Switzerland
>
> [log in to unmask]
> Tel. (41) 22 379 5293
> Fax (41) 22 379 5338
>
> On 1 juil. 2013, at 22:15, charlotte Riviere <[log in to unmask]>
> wrote:
>
>> Dear all,
>>
>> We had some problem in culturing DH1 strain lately in HL5 medium
>> (Formedium) : we have lots of yeast appearing after few days that we can
>> not get rid off.
>>
>> Is there any culture tricks to avoid such yeast contamination?
>> (The medium are autoclaved and filtered @0.22 µm)
>>
>> Best
>>
>> Charlotte
>>
>> --
>> Charlotte Riviere
>> Institut Lumière Matière (UMR5306 UCB Lyon 1 / CNRS)
>> Domaine Scientifique de la Doua
>> Bâtiment Léon Brillouin
>> 43 Boulevard du 11 novembre 1918
>> 69622 Villeurbanne France
>> Tél : +(33) (0)4 72 43 27 96 Fax : +(33) (0)4 72 43 26 48
>> [log in to unmask]
>
>
>
>
>
---------------------------------------------------------
Prof. Dr. Ralph Gräf
Universität Potsdam
Institut für Biochemie und Biologie
ZELLBIOLOGIE
Karl-Liebknecht-Strasse 24-25, Haus 26
14476 Potsdam
Germany
Tel.: +49-(0)331 9775520
FAX: +49-(0)331 9775522
http://www.bio.uni-potsdam.de/professuren/zellbiologie/aktivitaten
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