Oh the excitement, I actually didn’t know what to expect! I’ve been in labs before obviously, but this was a project of my own now! Where I would be the only student doing this and basically discovering new results for my supervisor!
So starting at 9am and ending at 6pm, I can honestly say it was a long day! Obviously I had a lunch break and everything, but still very long!
Thing is with these types of experiments, there is a lot of centrifuging and treatment that needs to be done in order to get the final, correct medium that can be finally analysed via the chromatography machine- FPLC: fast protein liquid chromatography. In our experiment we are basically only wanted to separate the different supercomplexes in the photosynthetic membrane; to mainly get the fraction of photosystem II, so we can further analyse the pigments in the photosystem II via HPLC: high performance protein liquid chromatography that are involved in light stress. Obviously, understanding the underlying mechanisms of the photosystem II will really aid in my future plans of further being a researcher in artificial photosynthesis J
Anyways here is photos to explain the day:
|This is a device which basically spins the spinach around in hand controlled speeds, hence drying washed spinach rapidly! We obtained the spinach fresh from Whitechapel market in the morning.|
|This is me chopping up the stems and midrib of the spinach preparing them for light treatment. I was initially quite slow as I didn't want to damage the spinach leaf and lose good material.|
|The spinach leaves ready for light treamtment.|
|The spinach leaves covers in cling film and starting light treatment.|
|Measuring the light intensity for the light treatment~ it was between 450-500 micro Eisenstein. This device is so cool it like an botanist stethoscope, haha!|
|Further measuring of light intensity and you can see the other plants grown by other researches at Queen Mary, in the plant room :)|
|Reading through the instructions and adding extra notes for next time.|
|While waiting for the centrifuge cycle, preparing of the buffers and mediums ready to add to the pellet of the centrifuge tubes.|
|The osmotic medium added after the break medium which breaks the photosynthesis membrane into smaller pieces, the break medium is added for 30 seconds, before the osmotic medium is added.|
|Centrifuge walkthrough, left to right- Time: 1.41 seconds remaining of the spin, Rotations per minute: 4000, Temperature: 4 degrees and not sure about what the 9 means, think its pre-set!|
|Taking buffer from the FPLC machine~which we need for further steps in the instructions.|
|Other picture of the FPLC machine, looks pretty cool~ overwhelming at first.|
So by the end of the first full day in lab I have completed pretty much nearly all the steps in the protocol plan for the 8 weeks! I was initially worried if I would have enough time in the 8 weeks to do it all, as I didn't know anything about how long each step will take. However, after today I understand the whole project alot more! That 8 weeks is plenty time to get really nice results in this project. That mainly the 8 weeks will be analysing the fractions that I obtain from solubilisation, to find the efficiency of Violaxanthin de-epoxidation in PSII supercomplex.
I am quite excited to move onto outline step B. I always think working with mutants is interesting as they can clearly indicate the purpose of certain complexes in each organisms.
Tomorrow I shall be analysing the results obtained from FPLC