Isolation and Culture of Bioluminescent Bacteria

Update: Keep your bacteria in the dark. I had been doing this without meaning to, and was confused to hear a fellow microbiologist growing a derivative of my cultures in her lab and getting no glow. When she grew them in the dark, they glowed again! Brief exposures have no effect. It’s the prevailing light conditions that seem to decide whether to stay active. This may be due to the light breaking down the little peptide the bacteria use to detect whether there are enough of them to glow meaningfully, or it might be a deliberate evolutionary adaptation. Who knows? :) I will update the document soon to add this consideration. Added pictures to this post, meanwhile. Finally got a good camera and took some.

In the logical follow-up to my last guide, which illustrated the production of homebrew bacterial media, here is a protocol for isolating bioluminescent bacteria from fresh seafood. The project was inspired by Mac Cowell’s successes. It is heavily based on the Indiana Biolabs protocol, differing in that it offers more background and suggests streaking your cells right away (I floundered for a while with mixed, impure cultures before finally streaking them out correctly on a petri dish and getting a pure culture).

It was written in a hurry, so if there are errors, omissions, or if it doesn’t make enough sense or explain things sufficiently, please let me know.

The document is available as a PDF and as an editable Openoffice.org document. It is released under a Creative Commons Attribution, Sharealike 3.0 license.

Again, please let me know if you use this protocol, and tell me how it worked out for you!

In related news, now might be a good time to point everyone in the direction of Applied Micro Systems, who sell lab equipment, supplies and mushroom cultivars for people interested in homebrew microbiology and mycology.

Some of my P.phosphoreum cells growing on tissue partially immersed in liquid broth. They glow brightest when grown like this or on a long agar slant:
Several repurposed jars full of bioluminescent bacteria