Team:Paris Liliane Bettencourt/Notebook/2011/09/09/

From 2011.igem.org

Team IGEM Paris 2011

Hovannes-Baptiste

Preparation of slides

Dilution of overnight cultures : 3610 (noted 3610 gfp-) and 3610 with GPF (noted 3610 gfp+) .

We waited to an OD of 0.6 (600 nm).

Two well slides: 1-control (3610 gfp-) 2-Mix (3610 gfp- and 3610 gfp+)

Observation

-37°C Microscopy-

We observed the plate with TRANS and GFP-filter settings on the Nikon microscope. We closed the fluorescence lamp diaphragm as much as we could without obscuring everything(1/16). The 3610 gfp+ strain proved to exhibit a strong fluorescence again. We had one experiment of approximately 5 hours. We took advantage of the automatisation of the Nikon to follow 5 different spots. However, the fine tuning of the Nikon seems to still escape us since we obtained only blurred image. We will retry with a better chosen focus tomorrow. Because of this blur and of a rapid bleaching of GFP, we can not use these images to conclude anything. Our preparation was also still not concentrated enough in the beginning. We are going to try to concentrate it even more in the next days.

3610 gfp-/3610 gfp+ : 37°C
3610 gfp- and 3610 gfp+ strains at 37°C t=0min position 1 (trans image)	3610 gfp- and 3610 gfp+ strains at 37°C t=0min position 1 (gfp image)
3610 gfp- and 3610 gfp+ strains at 37°C t=180min position 1 (trans image)	3610 gfp- and 3610 gfp+ strains at 37°C t=180min position 1 (gfp image, very blurry)