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Applications of STED fluorescence nanoscopy in unravelling nanoscale structure and dynamics of biological systems

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Abstract

Fluorescence microscopy, especially confocal microscopy, has revolutionized the field of biological imaging. Breaking the optical diffraction barrier of conventional light microscopy, through the advent of super-resolution microscopy, has ushered in the potential for a second revolution through unprecedented insight into nanoscale structure and dynamics in biological systems. Stimulated emission depletion (STED) microscopy is one such super-resolution microscopy technique which provides real-time enhanced-resolution imaging capabilities. In addition, it can be easily integrated with well-established fluorescence-based techniques such as fluorescence correlation spectroscopy (FCS) in order to capture the structure of cellular membranes at the nanoscale with high temporal resolution. In this review, we discuss the theory of STED and different modalities of operation in order to achieve the best resolution. Various applications of this technique in cell imaging, especially that of neuronal cell imaging, are discussed as well as examples of application of STED imaging in unravelling structure formation on biological membranes. Finally, we have discussed examples from some of our recent studies on nanoscale structure and dynamics of lipids in model membranes, due to interaction with proteins, as revealed by combination of STED and FCS techniques.

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Acknowledgements

The authors thank DST for funding through a special DST – IRHPA project. I.I.P also thank the IISER Pune – Leica Micro Imaging Center at Indian Institute of Science Education and Research, Pune, for STED imaging. Authors are also grateful to Prof. K. Ganapathy Ayappa and Prof. Sandhya S. Visweswariah, IISc, for productive discussions. We are grateful to Dr. Nirod Kumar Sarangi for his help in some experimental work.

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  1. Department of Physics, Indian Institute of Science, Bengaluru, 560 012, India

    C Roobala & J K Basu

  2. Center for BioSystems Science and Engineering, Indian Institute of Science, Bengaluru, 560 012, India

    I P Ilanila

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Roobala, C., Ilanila, I.P. & Basu, J.K. Applications of STED fluorescence nanoscopy in unravelling nanoscale structure and dynamics of biological systems. J Biosci 43, 471–484 (2018). https://doi.org/10.1007/s12038-018-9764-3

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