Abstract
Photolysis of caged compounds is a powerful tool for studying subcellular physiological functions. Here we describe protocols for the alignment and calibration of a focal uncaging system. We also report procedures for convenient quantitative calibration of uncaging. Using these methods, we can achieve submicron lateral resolution of photolysis and probe biological function in spines, the smallest signaling compartments of neurons. Initially, the entire alignment procedure takes 4–6 h to perform; periodic fine-tuning of the system takes 1–2 h.
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Acknowledgements
We thank G.M. Wittenberg and B. Kuhn for advice and discussion. This work was supported by the Rita Allen Foundation, the Whitehall Foundation, and National Institutes of Health grant NS045193. S.S.-H.W. is a W.M. Keck Foundation Distinguished Young Scholar. D.V.S. was supported by a Burroughs-Wellcome Interfaces of Science fellowship.
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Sarkisov, D., Wang, SH. Alignment and calibration of a focal neurotransmitter uncaging system. Nat Protoc 1, 828–832 (2006). https://doi.org/10.1038/nprot.2006.124
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DOI: https://doi.org/10.1038/nprot.2006.124
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