Notes (still writing on phone, sorry again for formatting.)
Yves Mely
FRET-FLIM visualize oligomerization
FCS measure N D and monomere dimer multimere. could we id twi dimer vs heterodimer?
measures kinetic rate constants for ligand receptor binding by id number and lifetime of each state.
how about with twist antibody? and two color labeling?
Elizabeth Rhoades
disordered protein spectroscopy.
lots of fcs curves gives fraction bound at different concrntrations of ligand.
Adaptive optics
many images much less than difraction limited due to diffraction distortion. Make a guide point back to a perfect diffraction limited spot by deformable mirror.
Joel Kubby
Adaptive optics for drosophila using fluorescent dextran beads as guide star.
peter Kner
Guess and check adaptable mirror, score sharpness. minimize extra images, reduce bleaching. Orthogonal measurements+ theoretical depth averaging- > fewer images.
fast focusing deformable mirrors. using MEMs. Compare to deconvolution- challenge is correct measure of scope pointspread fxn. Can use AO to. improve deconvolution
Na Ji (Janella Farms)
combining AO imaging with superresolution to get back diffraction limited spots from deep tissue.
Use liquid crystal spatial modulator instead of deformable mirror and use 2photon excitation. liquid crystal needs its own abberation correction.
brain imaging of bead 250 um in gives ghost images before correction.
getting away from beads. (could we use our diffraction limited STORM spots as beads).
correct using sample correlation (no need for beads).
field 100x100x100
apply to Ca imaging. Also works for confocal and superresolution widefield.
Jan Liphardt
distribution of processes matter ie restrain assembly mechanism.
Moerner
eYFP naturally photoswitchable / blinks.
also design small photo-activatable, targatable small dyes
3d traxking of single mRNA movement in 3d. 2d trcking projections don’t estimate well. Do 3d with helix transform.
A Pertsinidis
super-res live spatial tx reg.
subtract drift. .1C 100nm error. Better to actively stabilize while measuring distance. Use multi color dots as molecular ruler. 2color grid mapping.
challenge: inhomogenities in ccd detectors.
can we estimate how many transcripts per interaction by visualizing loops? Visualize histone modifications and correlate to transcription?
Thomas Derringer
super-res optical fluctuation imaging.
emitters fluctuate independently. correlation for 2 goes with 2 power, which is narrower tgan original gaussian.
constant background doesn’t fluctuate. Take movie analyze auto-correlation- don’t throw out the time data. don’t need to stochastically activate and bleach. need fluctuating signal.
readily do 3d.
how much fluctuation in emission do you need?