Single molecule methods with applications in living cells
•Knowledge about intracellular kinetics can be obtained without synchronization.•Single molecule sensitivity enables studies of processes involving low copy number species.•Recent development in open source software and instrumentation allows for straight forward implementation.
Our knowledge about dynamic processes in biological cells systems has been obtained roughly on two levels of detail; molecular level experiments with purified components in test tubes and system wide experiments with indirect readouts in living cells. However, with the development of single molecule methods for application in living cells, this partition has started to dissolve. It is now possible to perform detailed biophysical experiments at high temporal resolution and to directly observe processes at the level of molecules in living cells. In this review we present single molecule methods that can easily be implemented by readers interested to venture into this exciting and expanding field. We also review some recent studies where single molecule methods have been used successfully to answer biological questions as well as some of the most common pitfalls associated with these methods.
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Journal: Current Opinion in Biotechnology - Volume 24, Issue 4, August 2013, Pages 737–744