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Time-correlated single photon counting FLIM: some considerations for physiologists
Claire N Medine1, Angela McDonald, Axel Bergmann
1Centre for Integrative Physiology, University of Edinburgh, Hugh Robson Building, George Square, Edinburgh EH8 9XD, Scotland, United Kingdom.
Microscopy Research and Technique
|March 31, 2007
Summary
Cellular imaging now allows minimally invasive study of protein interactions in living cells. New techniques enable spatial resolution of protein-protein interactions and rates within cells, merging in vitro and in vivo approaches.
Area of Science:
- Cellular biology
- Biochemistry
- Biophysics
Background:
- Protein interactions are fundamental to cellular machinery.
- Traditional methods for studying protein interactions were primarily in vitro biochemical assays.
- Recent advances enable in vivo studies of protein interactions within living cells.
Purpose of the Study:
- To discuss recent developments in cellular imaging for studying protein interactions.
- To highlight the transition from in vitro to in vivo experimental approaches.
- To present findings relevant to physiologists studying protein dynamics.
Main Methods:
- Minimally invasive cellular imaging techniques.
- Biochemical approaches for protein interaction studies.
- Time-Correlated Single Photon Counting (TCSPC) imaging for interaction rates.
Main Results:
- Cellular imaging now permits the study of protein interactions in living cells.
- Spatial resolution of protein-protein interactions is achievable in vivo.
- TCSPC imaging advances allow determination of protein interaction rates with spatial resolution.
Conclusions:
- Experimental advances allow biochemical experiments to be performed inside living cells.
- The integration of imaging and biochemical approaches offers new insights into cellular machinery.
- Findings may be valuable for physiologists investigating cellular processes.
