. Author manuscript; available in PMC: 2012 Jul 24.

Published in final edited form as: Nat Rev Immunol. 2011 Jan;11(1):21–33. doi: 10.1038/nri2903

Table 1.

Applications of imaging techniques

Research question	Live-cell imaging	Fixed-cell imaging
Molecular structure	No	Crystallography, electron microscopy
Conformational changes	FRET, single-molecule FRET	Crystallography, electron microscopy
Mobility of bound species	FRAP, FCCS, SPT	No
Intracellular activity of proteins	FRET sensors, FRET	No
Intracellular localization	Confocal microscopy, STED microscopy	Confocal microscopy, STED microscopy, SIM, PALM
Aggregation state of receptors	Anisotropy, FRET, PALM, STORM, FCCS and related analyses of molecular brightness	TEM, PALM, STORM, FRET
Mobility at the plasma membrane	TIRF microscopy, FRAP, SPT and sptPALM, confocal microscopy, STED microscopy	No
Cell morphology	Confocal microscopy, epifluorescence microscopy, TIRF microscopy, DIC microscopy	Confocal microscopy, epifluorescence microscopy, TIRF microscopy, DIC microscopy
Cell adherence to a surface	TIRF microscopy, DIC microscopy, IRM	TIRF microscopy, DIC microscopy, IRM, TEM

DIC, differential interference contrast; FCCS, fluorescence cross-correlation spectroscopy; FRAP, fluorescence recovery after photobleaching; FRET, fluorescent resonance energy transfer; IRM, interference reflection microscopy; PALM, photoactivated localization microscopy; SIM, structured illumination microscopy; SPT, single-particle tracking; STED, stimulated emission depletion; STORM, stochastic optical reconstruction microscopy; TEM, transmission electron microscopy; TIRF, total internal reflection fluorescence.