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Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy
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Watching cellular machinery in action, one molecule at a time
Enrico Monachino1,2, Lisanne M Spenkelink1,2, Antoine M van Oijen3
1Centre for Medical and Molecular Bioscience, Illawarra Health and Medical Research Institute and University of Wollongong, New South Wales 2522, Australia.
The Journal of Cell Biology
|December 17, 2016
Summary
Single-molecule techniques reveal the dynamic, stochastic behavior of biomolecules and complex protein systems. These advanced methods offer unprecedented insights into cellular processes like DNA replication, both in vitro and in vivo.
Area of Science:
- Biophysics
- Molecular Biology
- Cellular Dynamics
Background:
- Single-molecule manipulation and imaging are crucial for understanding biomolecular mechanisms.
- These methods overcome limitations of ensemble averaging, providing access to individual molecule dynamics.
- Recent expansion includes studying complex multiprotein systems and intracellular molecular behavior.
Purpose of the Study:
- To review force- and fluorescence-based single-molecule methods.
- To highlight applications in cytoskeletal motors and DNA replication.
- To discuss enhanced understanding of complex multiprotein systems' dynamics.
Main Methods:
- Force-based single-molecule manipulation (e.g., optical tweezers).
- Fluorescence-based single-molecule imaging (e.g., FRET, TIRF microscopy).
- In vitro and in vivo experimental approaches.
Main Results:
- Single-molecule approaches provide unique mechanistic insights into cellular processes.
- Detailed characterization of individual biomolecule behavior inside living cells is now possible.
- Complex multiprotein systems exhibit highly stochastic, non-linear dynamics.
Conclusions:
- Single-molecule techniques have revolutionized the study of biomolecular dynamics.
- Understanding protein complex behavior requires moving beyond deterministic models.
- Continued development of single-molecule tools is vital for elucidating complex biological systems.
