Simultaneous widefield single molecule orientation and FRET microscopy in cells.
We combine single molecule fluorescence orientation imaging with single-pair fluorescence resonance energy transfer microscopy, using a total internal reflection microscope. We show how angles and FRET efficiencies can be determined for membrane proteins at the single molecule level and provide data from the epidermal growth factor receptor system in cells.
Webb
S E D
SE
Science and Technology Facilities Council, Daresbury Laboratory, Warrington WA4 4AD, UK. stephen.webb@stfc.ac.uk
Rolfe
D J
DJ
Needham
S R
SR
Roberts
S K
SK
Clarke
D T
DT
McLachlan
C I
CI
Hobson
M P
MP
Martin-Fernandez
M L
ML
D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D003461:Crystallography / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D066246:ErbB Receptors / Q000378:metabolism* / Q000648:ultrastructure*; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D006801:Humans; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D011487:Protein Conformation
D066246:ErbB Receptors