I am using monoolein as the lipid and I have generated several interesting results from the 96 screening conditions used, but I cannot find any images anywhere that are the same as mine. A lot of the conditions produce rivulets that contain small sparsely spread blue dots. Over a period of a week, these blue dots accumulate and cause a striation pattern in the rivulet. Can anyone tell me what is going on?
Any chance you can post a link to your picture? This was the first trouble I had when I started out, interpretation was pretty hard and I had to resort to poking it etc. to find out.
By the end, I'd resorted to a scheme of first finding out if it was protein or salt, then finding out whether it was seedable, etc. but there was always a little bit of art mixed with this science.
I'd look to see if there's any source for the blue dots in any of your buffers/conditions, and see how solid they are.
Hi Kannan
Thanks for your interest. I have attached a file with several of the pics. I've included the conditions they stemmed from.
Sorry, was away on vacation and was hoping more experienced people might chime in! I'm no final expert, but in your position I would screen conditions around everything except the first one.
The first seems to be stuck in that frustrating and ambiguous "birefringent precipitate" or "phase separation" purgatory, which might be great if you have nothing else going for you, but the others seem to be very apparent as crystalline and what I'd aim for - especially the one that crystallized for 3 days and then resolubilized.
I usually like to check how reproducible the crystals are before playing with them and finding if they're protein crystal. Good luck, you seem to be a lot closer than I would've guessed from your initial post!
Thanks for taking the time to look at them Kannan. Much appreciated.
Hi - your crystallisation is going from cubic phase to a highly birefringent lamellar phase - its difficult to see any crystals in this phase ;\ you often get it when your salt or peg concentrations are too high (in your case the salt is way too high to support cubic phase) - its a good idea if you're using commercial screens that are not designed for lcp to dilute them - to 50-70%. hope this helps! sorry i don't log onto RG very often....
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