Hey there! Hope you're getting on okay?

How's this for you:

VWR product code 391-1261 or 391-1262

Thanks,

Elyse.

Hi,

I am thanks, half way through my PhD now. Hope all is good at Chester.

I think I am going to try a few seals and test them with resazurin and see which can keep the oxygen out.

Cheers,

Elizabeth

Hi Elizabeth,

As an alternative you could also try regular PCR seals from BioRad (for sealing 96 well plates in thermocyclers). It has worked well in our lab.

Cheers.

András

Thanks very for the advice. I will try them.

No answer here, sorry. Just curious, Elizabeth, if you have made any progress on this front.

I'm interested in looking at anaerobic growth in 96 well plates, and our plate reader isn't in a glovebox.

Andras, just want to clarify: Were you suggesting that the seals would keep a 96 well plate anaerobic outside of a glovebox??

Thanks!

I am also interested in growing some anaerobic environmental isolates in 96-well format on different substrates, then screening OD using a microplate reader (outside of chamber). Has anyone tester any particular sealing films that stay anaerobic outside of the chamber

Thanks!

Did anybody get this working - am thinking of doing something similar?

I actually ended up trying a number of different brands and they all worked when sealed in anaerobic cabinet before removing. I would recommend just trying whatever you have to hand or which can be used for other applications like PCR plate seals

Have you tried a normal 96 well plate and wrap the gap between the lid and the base with parafilm inside the anaerobic cabinet? If so, how well does it work compared to the PCR seal?

It works in terms of keeping the culture anaerobic but it wouldn't fit in our micro-plate reader. 

I'm planning to use sealing film as well since my micro-plate reader doesn't accommodate my plates with the lid. Elizabeth, does the film interfere with the OD reading? I'm reading yeast growth at OD600

I don't believe so. Construct your blanks in the same plate with the seal. 

Hi Elizabeth, Axygen covers these seals. The sku is BF-400

 On this question....what 96 well plates do you recommend to you for experiments like this?  So many of them seem to be oxygen permeable, and I want to prepare anaerobically then do serial readings in a plate reader in an open lab?

I've successfully used these ThermaSeal PCR sealing films (Excel Scientific) to provide an anaerobic environment for the culture of green sulfur bacteria, however the growth on the outer wells is unreliable because of the texturing on the edge of my 24/96 well plates makes them difficult to seal. Stick to the inner wells (Rows B-G, columns 2-11) if you want to have more consistent results.

The plastic on these ThermaSeal tubes is polypropylene, which is OK as an oxygen barrier for some anaerobic strains such, as green sulfur bacteria, which can scavenge oxygen, allowing them to survive in its presence, but not grow. Other bacteria may flat out die. If that's the case, you might want to try to explore a plastic film material that's designed to prevent gas exchange, such as Ethylene Vinyl Alcohol (EVOH). This plastic is used widely in the food industry to keep oxygen out when packaging (according to Wikipedia), so it may be very useful to grow cells with However, I don't think anyone sells this film in a form that makes them convenient for use with 96-well plates. Perhaps with a heat sealer you can make an anaerobic seal around the plate.

In my old lab, I would carefully melt birthday candles (a cheap source of paraffin that's conveniently shaped) to make glass-glass seals for the anaerobic growth of photosynthetic bacteria between a microscope slide and a cover slip. It may be feasible to just seal a 96-well plate with paraffin (or a combination of paraffin with petroleum jelly to make the paraffin more maleable). Nail enamel also works really well, but it can be toxic to your bacteria if it comes in contact with the growing medium and is generally more "runny" than paraffin, so it may not work well in sealing larger gaps.