The Xanthophyll Cycle in Photosystem II

Thursday, 5 June 2014

UK’s Beautiful Sunshine Rays + First Run of HPLC

Today was my first time on the Fogg's rooftop (Fogg Building= Queen Mary’s School of Biological and Chemical Sciences building). It’s basically where all the greenhouses are for the school. Here are some lovely pictures of it:
Here are the several green houses on the rooftop. If you can see the blue door in the picture, there are more on the other side of the door!

Green house shot 2# : As you can tell the green houses are quited dated and I believe money should be spent on making them nicer! Apparently when a glass window falls or breaks, inevitable when the greenhouse is that old, it costs so much to replace just the glass, so why not replace it all? Guess with most thing its about funding! Currently Fogg building is undergoing alot of work inside with refurbishing and building new offices~ so guess that's where the money is going !

Loving the clouds in this picture, and you can just tell by the building that this is iconic East London!

East London view~

A photo of the rooftop, East London views and the pathway leading back to the fifth floor of the fogg building with the ladder steps. As you can tell the Fogg building is very peculiar in itself for the fact it has this Lego theme going on it. I'll blog about the architecture of the actual Fogg building some other blog post! So look out for it in the near future!!!

Better view of what I described the photo above!

Petra (Ruban Labs technician) giving me a tour around Ruban Lab's Greenhouse. Ohhh looking at this picture reminds me of how amazing the weather was that day! ^^

Botanist playarea! Haha

Anna's Plant. Anna is a final year Phd student of Ruban's. She works on hybrid plants! Frankenstein like she says it!

Healthy plant growth

They look like they are dying but this is how we obtain the tiny seeds from them! Where Petra hand is positioned is basically where all the fine seeds are located.

We started off the lab day discussing about the fine details from my previous meeting with Professor Ruban and how we can further change the parameters to improve our solubilisation for PSII complexes. After this, we headed back down to the fourth floor, where Ruban's Lab is.

Finally the day had came and it was HPLC time! I’ve never ever used a HPLC machine in my life! Since the 3 weeks of being here I've always anticipated that we would eventually use this to further analyse certain fractions obtained from FPLC. I’ve always wondered how this machine works, how we would use it! New experiences are always the most exciting.

I must say, I really do prefer the HPLC machine over to the FPLC. I find HPLC machine more compact and nicely in place. The FPLC for me feels abit large and awkward in the sense you have the fraction collector part of FPLC. To be honest, maybe it is the positioning of FPLC, as it is placed awkwardly next to a beam that makes it so awkward to use, especially when you are trying to inject your sample from a syringe into the injection valve.

Anyways I'll leave it up to you guys to decide what you think of each, here some pictures!:

Do you see what I mean? The fractions are nicely placed in this circular spinning part of HPLC, everything is compact and nicely in placed! If you go back to my blog post about FPLC you can see the fraction collection part of the machine isn't as nicely compacted as HPLCs!

This is Petra adjusting the HPLC machine, checking if everything is set properly. Above of the machine we have open the flap , where we can see the buffer wires. We can observe if the machine is working properly if there are no bubbles in the buffer wires and if the buffers are actually moving through the tube!

Our very concentrated A8 fraction sample of PSII. From our second run of FPLC and isolation of stacked thylakoids.

Labelled vials for the different isolated PSII fractions from different experiments.  The number 1's are from the 1st isolation of stacked thylakoids. 1c being the most concentrated, 1b being a 1:1.25 dilution and 1a being a 1:2 dilution. The number 2's are taken from the 2nd isolation of stacked thylakoid experiment. C being the most concentrated going to slowly up to a lower dilution factor.  2 labelled vials are non acidified samples and, 3 labelled vials are acidified sample.

All the vials nicely placed in, if you see to the back of the machine there is basically a needle attached to the metal moving thing. This is the needle which injects into the vial and samples for the HPLC run.

Side view of HPLC

A better picture of all the vials in place with their caps on. In the middle of the cap is a circular paper disc which the needle can easily pierce.

Computer part of the HPLC. Yes you can clearly see windows 2000! You might be thinking why is this not windows 8?! Or something more up to date? Basically most of these chromotography or any lab machines were built a while back and obviously configured to work with windows 2000 only. For Ruban's lab to change and update everything to windows 8 could take a process of an entire month. So if it works then its fine. Despite the fact it is awfully slow!!!! -.-

With the lid on!

Lovely windows 2000 again! Did i mention it was windows 2000 professional?! :O

Liquid nitrogen, being daring and placing my hand on the vapor ! Basically its fine as when the vapour touches your skin it immediately evaporates! Don't ever actually immerse your entire hand in the liquid nitrogen its equivalent to putting your hand in burning oil, besides your hand would freeze in pain. For me I think freezing to pain is worst as I am so sensitive to cold. I remember doing a lab experiment to see how tolerant I was to placing my hand in icy waters, yeah I didn't last long....*roll eyes* :)

The program that processes the data collected and analysed by HPLC. A nice mouse mat which probably came with the software! How kind of Chromeleon!

Chromeleon on the desktop loading!

Literally, this is five minutes after and yet it still fails to load.... well it did load eventually but obviously windows 2000 is old, it's gonna be slow!

Setting the parameters for our HPLC run~ oooohhh colours!!!! You might wonder how on earth would I know how to use this as a undergrad, wouldn't I get confused? Basically we have this HPLC for dummies walk through which I just follow the instructions to~ it is slightly overwhelming, but if you can read you'll be fine :P

To conclude, this was a very interesting day. The HPLC run was basically a test run as our fraction samples did not have 100% PSII complexes only, they were not fully unsolublised. Today was basically for me to see how HPLC works and to get use to the machine itself :)

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