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Correlating Scanning Ion Conductance and Super-Resolved Fluorescence Microscopy

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Scanning Ion Conductance Microscopy

Part of the book series: Bioanalytical Reviews ((BIOREV,volume 3))

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Abstract

The development of diffraction-unlimited fluorescence microscopy has paved the way for a detailed understanding of cellular dynamics since live-cell investigations with resolutions below 100 nm have become available. This allows studying labelled specimen such as proteins and their distribution within a cell at a new level of detail. However, examining the cell membrane as a site of interaction between the cell’s internal processes and external signals with fluorescence microscopy (FM) leads to the difficulty that certain types of fluorescent dyes can alter the physicochemical properties of a cell. Investigating the cell membrane in a contact-free way and without any labelling or special pre-treatment is one of the greatest strengths of scanning ion conductance microscopy (SICM), which uses a fine pipette tip to determine the specimen’s topography. Since FM and SICM can both be applied to living cells and provide a resolution below the diffraction limit, correlating data from these two techniques seems to be a promising approach to further the understanding of the cell membrane’s organization and the mechanisms involved in transport processes across it. In the following, we briefly introduce the different approaches to super-resolved fluorescence microscopy (SRFM) and review applications of correlating SICM and FM (diffraction-limited) as well as SICM and SRFM (diffraction-unlimited). Finally, we discuss potential implementations of combined SRFM/SICM instruments and pitfalls that may arise during the development of such instruments.

Patrick Happel, (deceased), Happel wrote this chapter while at Nanoscopy Group, RUBION, Ruhr-Universitðt Bochum, Bochum, Germany

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Acknowledgements

The authors acknowledge support from the Volkswagen Foundation (grant 88 390) and the German Research Foundation (grant 411517989).

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Authors and Affiliations

  1. Nanoscopy Group, RUBION, Ruhr-Universität Bochum, Bochum, Germany

    Patrick Happel

  2. Nanoscopy Group, RUBION, Ruhr-Universität Bochum, Bochum, Germany

    Annika Haak & Astrid Gesper

  3. Institute for Physiological Chemistry, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany

    Astrid Gesper

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  1. Institute of Applied Physics, University of Tübingen, Tübingen, Germany

    Tilman E. Schäffer

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Happel, P., Haak, A., Gesper, A. (2022). Correlating Scanning Ion Conductance and Super-Resolved Fluorescence Microscopy. In: Schäffer, T.E. (eds) Scanning Ion Conductance Microscopy. Bioanalytical Reviews, vol 3. Springer, Cham. https://doi.org/10.1007/11663_2022_13

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