User:Linh N Le/Notebook/2009/06/11: Difference between revisions
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*Right now, brigette is looking at the 3ul aa soln | *Right now, brigette is looking at the 3ul aa soln | ||
**They are still photobleaching quite fast, so it is really hard to tell if the tubes are breaking apart or not | **They are still photobleaching quite fast, so it is really hard to tell if the tubes are breaking apart or not | ||
*Now trying it with the stock of AF that brigette made last week | |||
*Doing | |||
#10ul of the tubes + 3ul acetic acid | |||
#2.5ul of AF | |||
#1ul of 2M dextrose | |||
#80ul of BRB80T | |||
*took the soln above and gave briggete and sample to look at | |||
*took the soln above, added 3ul more aa to the whole stock, make sample for brigette to look at | |||
*note: I only used maybe 1/10 of the AF cocktail today | |||
**refroze the aliquot, to save money, marked the aliquot with "thawed 6/11" | |||
**I hope AF is ok with being thawed, refrozen and thawed again | |||
*Brigette and I have spent the good part of an hour trying to image our new samples. | |||
**~45mins after creation, I took a look a new sample of the AF+MT(3ulaa)+dex+brb80t+2ulaa mix and saw some floaties. | |||
**The AF didnt seem to work very well. The background went black too fast! | |||
*Getting frustrated at the fact we could not see anything, I went ahead and sampled out the pure tubes +1.5ul aa | |||
**We saw plenty of tubes, and even though they do fade quite rapidly, you can still see them faintly in the background | |||
*Conclusion of the day: AF does not screw up MT. AA does not screw up MT. Mixing the 2 messes everything up! | |||
*In a last ditch effort, Brigette added 10 more ul of MT(with 3ul of AA) to the AF mixture | |||
**Hopefully more MT's allows us to see some | |||
*On a side note: we really need another microscope as Andy came back into the lab and wants to work on the tweezers, but we are still looking at samples! | |||
*MT's, no antifade, with aa, can see tubes. MTs with AF, with aa, still fades badly, but not tubes. "bad" AF that doesnt stop fading, +MTs, can see tubes, but they fade fast (as we saw last week). MTs + aa +AF (whether or not its good b/c it was in the freezer, or even if I messed it up by not thawing it right) no tubes. aa+af=bad for tubes. | |||
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