I’m back in Leiden, with the aim of putting together the modified fab@home 3D printer and the plates of bacteria in the lab. The following 2 weeks will be the moment of truth, and thus we’ll see if he project was worthwhile.
A lot of preparation has already been kindly done by Maarten de Smit, who has arranged for the production of large quantities of LB agar, with 0.75% agar content (this is the agar content ratio of what is called the ‘top agar’). Trudie Bowers has provided me with all sorts of equipment and materials at the lab, like arabinose, liquid LB, pipettes, Erlenmayer flasks, liquid water bath, gas torch, large pipettes with rubber pumps, etc. Anne Kienhuis is organizing everything for me very kindly.
I will be working with 12x12cm plates to create my images.
The daily routine here will consist of maintaining a stable supply of plates full of bacteria in LB agar, so I can put them into the printer, and then experiment with the plates in the printer. To provide a stable supply, Maarten has grown some bacteria, taken from the cryogenic storage and spread out on a plate. Using this bacteria, overnight liquid cultures have to be made each day. Using these liquid cultures, the plates can be made themselves with bacteria, which also take overnight, and have to be put into an incubator.
Today I made a liquid culture with 100ml of liquid LB and some ampicillin. Here I’m in a bit of a trouble, as the ampicillin bottle says 250mg/ml, which seems an extremely high concentration, about 5000 times more concentrated than the 50μg/ml carbenicillin I used to use. Thus I put 1μl of it into 100ml of liquid LB, which should be 250μg of ampicillin. And of course a scrape of bacteria from the plate with the spread out bacteria.
I also made 10 plates, 12x12cm each with LB agar and liquid cultures. Used 1 liter of LB agar (0.75% agar), and put 50ml of liquid culture into it. I had the same dilemma here with the ampicillin, so these guys also got only 1μl, which should be 250μg again. I wonder if it works.
Afterwards, I started to fire up the printer and print on plates prepared by Maarten. At first of course there were some problems, but at the end it seemed to work out. Well, we don’t know of course – I got reasonably small drops of arabinose, but we’ll have to see tomorrow if they work out. Now I’m using a 16×16 pixel grid, which is old-school icon size.