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Movies

Much of our work involves fluorescence imaging of living cells. Below are some movies showing our probes and reporters at work in living cells.

 
  (QuickTime) Movie showing the movements of single AMPA receptor molecules, labeled with quantum dots (red), on the surface of living hippocampal neurons. The neurons are also expressing a synaptic marker, PSD95-YFP, shown in green. The movie is 3-fold faster than real time. From Howarth et al. PNAS 2005.
   
  (Avi) Movie showing the response of a FRET reporter of histone phosphorylation (specific for serine 28 on histone 3) in living HeLa cells undergoing cell division. Red signifies high FRET (and high phosphorylation levels), blue signifies low FRET (and low phosphorylation levels), and green is intermediate. The reporter displays a rapid increase in FRET 5-15 minutes after breakdown of the nuclear envelope. High FRET is sustained for several hours and then drops slowly after cytokinesis (separation of the cytoplasm into daughter cells). Over 26 hours. From Lin et al. Angew. Chemie 2004.
   
(Avi) Movie showing the response of a FRET reporter of Abl tyrosine kinase activity in a 3T3 fibroblast stimulated with PDGF. Red signifies high FRET (and high Abl activity), and blue signifies low FRET (and low Abl activity). Upon stimulation with PDGF, reporter phosphorylation increases in the cytosol (yellow to red pseudocolors), while remaining unchanged in the nucleus (blue). Reporter phosphorylation is greatest in the membrane ruffles (bright red) along the lower right edge of the cell. Over 50 minutes. From Ting et al. PNAS 2001.
   
(Avi) Movie showing the response of a FRET reporter of EGF receptor activity in a B82 mouse fibroblast stimulated with EGF. Red signifies high FRET (and high EGF receptor activity), and blue signifies low FRET (and low EGF receptor activity). Upon addition of EGF, FRET first increases at the extremities of the cell, where the ratio of plasma membrane surface area to cytosolic volume is highest. The FRET change then propagates through the cytosol towards the center of the cell, while the nucleus remains relatively unchanged. This pattern is expected for a kinase that is localized at the plasma membrane and that phosphorylates the fluorescent reporter at rates comparable to the diffusion of the latter through the cytosol. Over 3 minutes. From Ting et al. PNAS 2001.
   

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