The recent advances in optical microscopy enable the simultaneous visualization of thousands of structural and signaling molecules as they dynamically rearrange within living cells. Super-resolution microscopy offers an unprecedented opportunity to define the molecular mechanisms of nanosensing through direct observation of protein movement. This technology provides a real-time readout of how genetically targeted molecular perturbations affect protein interactions. As we strive to meet the challenge offered by the opportunity to ask questions about the mechanism of cell that we never thought we could answer, we need to be aware that the new technologies are still evolving. The current limitations of each technique need to be considered when matching them to specific biological questions. In this review, we briefly describe the principles of super-resolution optical microscopy and focus on comparing the characteristics of each technique that are important for their use in studying nanosensing in the cellular microenvironment. WIREs Nanomed Nanobiotechnol 2011 3 247–255 DOI: 10.1002/wnan.130
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