Yesterdays experiment with growing Photobacterium Phosphoreum as a liquid culture came to fruition today. It seems that the ‘botched’ liquid culture from two days ago came to light today as well. The following images show the result, in a pitch dark room with some reflection off the wall (well, a tiled bathroom), at 10sec, f/4, ISO 800:
In short, I prepared two 100ml cultures (one PP BH and one PP LumA) in a 500ml Erlenmayer flask each, with the following composition each:
- Nutrient Broth: 8g/1000ml -> 0.8g
- NaCl: 30g/1000ml -> 3g
- Glycerol: 10g/1000ml -> 1g
- CaCO3: 5g/1000ml -> 0.5g
The ‘botched’ bottle from two days ago had the same ingredients, but in cca. 180ml of water. The result is well visible in a dimly lit room. I personally can’t tell the difference between the PP HB and the PP LumA variants.
The bacteria light up only when you oxygenize the culture, for example, you shake / stir the liquid culture. after that, it stays visibly bright for about 7 minutes. See the following image sequence, taken in a loosely lit room in 1 minute increments, at 3s, f/4, ISO 800:
Yesterday I did some experiments with Photbacterium Phosphoreum, received kindly from Simon Park, I received two different batches, one simply labeled PP BH, the other PP LumA. I grew out two of each on agar plates, with the following ingredients for 4 plates, 80ml of Agar total:
- Nutrient Broth: 8g/1000ml -> 0.64g
- NaCl: 30g/1000ml -> 2.4g
- Glycerol: 10g/1000ml -> 0.8g
- CaCO3: 5g/1000ml -> 0.4g
- Agar: 15g/1000ml -> 1.2g
I also tried to create a liquid culture, where I botched the ratios and ended up with twice the liquid for the amount of ingredients.
The plates lit up overnight, the liquid culture didn’t. They are quite bright when compared to Vibrio Fischeri. Below are images with various ambient light and photo settings used to take them. other than cropping the first two images, no modification has been made. The two plates on the right left are the PP HB plates, the right ones are the PP LumA plates.
last Thursday we performed the first set of DNA transformations with a small circle of friends, Ferenc Szalai, Attila Nemes and András Márton Juhász. this was a basic test of the lab to prove if multiple people can work in it together.
Yesterday I performed the first successful DNA transformation in the new lab, set up at FabLab Budapest. The transformation was simply the pGLO gene earlier isolated at the University of Szeged, into e.coli HB 101. The green spots on the image above are the bacteria with the Green Fluorescent Protein in then.
This gives reassurance that the crude tools here in the lab can do real work
Tomorrow, I’ll hold a presentation on bio.display at the Iwasaki Lab of the Center for Advanced Biomedical Sciences at Waseda University in Tokyo, at 2:30pm local time. See Iwasaki Sensei’s page on what he’s mostly up to.