Abstract
The most frequently encountered mechanisms and techniques in fluorescence microscopy are detailed in chapter 7. This includes time-domain fluorescence life-time imaging and its advantage over intensity based fluorescence imaging. This is followed by the description of the resonance energy transfer mechanism for a wide range of molecular pairs (Acceptor-Donor pair) that is central to many biological studies. Next, we discuss the generation of second and higher harmonics using a non-linear crystal for imaging applications. In the final section, fluorescence correlation spectroscopy and its variants are discussed. This enables study of numerous biophysical phenomena including, fluorescence fluctuation and diffusion in the focal volume.
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Appendix: Recommended Reading
Appendix: Recommended Reading
1.1 Books
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1.
Rigler, R. and Elson, E.S., Fluorescence Correlation Spectroscopy: Theory and Applications, Springer Series in Chemical Physics, Volume 65, 2001.
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2.
Diaspro, A. (Ed.), Optical Fluorescence Microscopy, Springer, 2011.
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Diaspro, A. (Ed.), Nanoscopy and Multidimensional Optical Fluorescence Microscopy, CRC Press, 2010.
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Hawkes, P.W. (Ed.), Advances in Imaging and Electron Microscopy, Elsevier, 2013.
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Novotny, L. and Hecht, B. Physics of Nano-Optics, Cambridge University Press, 2006.
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Selvin, P.R. and Ha, T. Single-Molecule Techniques: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2008.
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Mondal, P.P., Diaspro, A. (2014). General Fluorescence Imaging Techniques. In: Fundamentals of Fluorescence Microscopy. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-7545-9_7
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