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Freshness of a culture for staining purposes - (Dec/11/2007 )

I wish to do Gram staining and endospore staining and was wondering how fresh a culture should be.

Do I culture on agar or broth for this purpose?

I have a few suggestion using other media but I'll be using LB (agar and broth).
Another question is that since we know endospore formation in a bacteria is triggered by a not-so-good environment, won't a fresh culture means a good nutritional environment hence absence of spores?

Thanks people.


i'm a bit out of my league here (more histo than micro) but i'm thinking it depends on your definition of fresh and what you want from it.
ie - will there be breakdown of the culture and production of substances that you wouldn't find in a fresh culture - do you want them?
then its a case of balance - grams very easy to do, you could try a series over time to give you more of an idea.

not much of an answer i admit

hope it works out