Vitamins are for wusses (#arseniclife)

The GFAJ-1 cells I streaked on a plate of my less-than-specified culture medium are growing, and much faster than I had dared hope.  After less than 48 hr I can already see tiny (very tiny) colonies.  I can't estimate growth rate because I don't know how many cells are the colony,

I was worried that they might not grow at all, because this medium doesn't include the trace-element mix that Wolfe-Simon et al. added to their medium.  Instead I simply made up the medium with lovely Vancouver tap water.  It also doesn't include most of the vitamins they added - I only had stocks of thiamine and pantothenic acid, so they were the only vitamins I added.

I'm still considering solutions to the cell-counting problem.  Flow cytometry of acridine-orange-stained cells won't work very well because some of the cells are in small clumps of 2-10 cells.  Collecting known volumes of cells onto black filter membranes would probably work fine but would be a pain and use a lot of the filters.  I think the simplest solution is to make a 2-X acridine orange stock that contains a known density of 2 µ polystyrene beads, mix this with an equal volume of culture, and then count both the beads and the cells in random microscope fields-of-view.  I'm going to test this now.