Understanding biological dynamics: following cells and molecules to track functions and mechanisms
The dissection of the molecular circuitries at the base of cell life and the identification of their abnormal transformation during carcinogenesis rely on the characterization of biological phenotypes generated by targeted overexpression or deletion of gene products through genetic manipulation. Fluorescence microscopy provides a wide variety of tools to monitor cell life with minimal perturbations. The observation of living cells requires the selection of a correct balance between temporal, spatial and “statistical” resolution according to the process to be analyzed. In the following paper ad hoc developed optical tools for dynamical tracking from cellular to molecular resolution will be presented. Particular emphasis will be devoted to discuss how to exploit light–matter interaction to selectively target specific molecular species, understanding the relationships between their intracellular compartmentalization and function.
KeywordsFluorescence microscopy Cell tracking Fluorescence recovery after photobleaching Photoactivation Two-photon microscopy
This work was supported by IIT (Italian Institute of Technology, Genoa, Italy) and by IFOM (FIRC Institute of Molecular Oncology, Milan, Italy). The authors are grateful to Dr. Francesca Ballarini for her help in manuscript preparation.
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