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I am working with an undergraduate on a circadian experiment with crickets, where one of the things we're trying to sort out is whether the stable isotope tracers we plan to inject actually represent tracers and not a large nutrient bolus.

She's been spinning her wheels for the last couple of months for multiple different reasons. A large headache returned two days ago, when she started having problems again with hemolymph sampling. The ideal thing to do would be to be able to take a hemolymph sample, put it into some sort of solution to dilute it, and then split it and measure total protein, carbohydrates, and lipids on the subsamples.

This is not so easy to do when the hemolymph sample keeps coagulating.

The internets suggested trying to add ascorbic acid, and that seemed to help, but then we discovered that it's incompatible with the protein assay at any concentration.

In the meantime, I'm working on figuring out what I need to know to start measuring enzyme activities for five major enzymes of glycolysis and the TCA cycle and beyond (hexokinase, pyruvate kinase, glucose-6-phosphate dehydrogenase, citrate synthase, and cytochrome c oxidase). There's a lot to figure out, starting with what sorts of homogenization methods I'll need to apply, because of course they're going to differ depending on the enzyme and where it's found. Something tells me that cytochrome c oxidase is going to be the worst.

This is a somewhat unsatisfying stage of things, because I have to do a bunch of looking around and shopping for things without quite knowing what's going to pan out and what I'm going to screw up. The head cold doesn't help, either. I could stay home and try to rest, but I don't think that will do me a tremendous amount of good.



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October 2018


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