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Optogenetic Tools in the Microscopy of Cardiac Excitation-Contraction Coupling

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Microscopy of the Heart

Abstract

Microscopy became a scientific investigation method in the seventeenth century with the application of the first build microscopes on biological samples [1, 2]. Soon it became a popular method to stain samples in order to visualise particular (cellular and subcellular) structures [3]. These stains, either based on absorption or fluorescence, have limitations in respect to their specificity and are often toxic to cells, which limits investigations to short intervals or even dead samples. In 1987 the idea came up to use a fluorescent protein that was discovered 25 years before [4], in particular a green fluorescent protein (GFP) form the medusa Aequorea victoria to label cells and cellular structures [5]. With the sequencing and cloning of GFP, a so-called ‘green revolution’ started, which led to regular usage of fluorescent proteins as markers or sensors (for details see below) in the majority of cellular research in physiology, microbiology, pharmacology, molecular biology, anatomy, cell biology, biophysics and many other biomedical fields. Although the expression of the fluorescent proteins and their optical investigation can already be regarded as optogenetic tools, this term was only applied when the optical properties of proteins were used to manipulate cells. The best-known example of such a protein is the channelrhodopsin, a light-gated ion channel [6, 7]. When this ion channel is expressed in a membrane and illuminated with light of the appropriate wavelength, the channel will be activated and opened, which results in passive transportation of ions across the membrane and a change of the membrane potential. However, within this chapter we consider both aspects, the observation and the manipulation as optogenetic tools. To use the optogenetic tool, the genes of these proteins need to be transferred into the cells to allow the expression of the protein. For an overview of gene delivery into target cells, see [8].

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Authors and Affiliations

  1. Theoretical Medicine and Biosciences, Saarland University, Homburg/Saar, Germany

    Lars Kaestner & Qinghai Tian

  2. Experimental Physics, Saarland University, Saarbrücken, Germany

    Lars Kaestner

  3. Institute for Neurophysiology, Hannover Medical School, Hannover, Germany

    André Zeug

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  1. Lars Kaestner
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  2. André Zeug
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Editors and Affiliations

  1. Theoretical Medicine and Biosciences, Saarland University, Homburg/Saar, Germany

    Lars Kaestner

  2. Preclinical Center for Molecular Signal Processing, Building 61, Saarland University, Homburg/Saar, Germany

    Peter Lipp

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Kaestner, L., Zeug, A., Tian, Q. (2018). Optogenetic Tools in the Microscopy of Cardiac Excitation-Contraction Coupling. In: Kaestner, L., Lipp, P. (eds) Microscopy of the Heart. Springer, Cham. https://doi.org/10.1007/978-3-319-95304-5_5

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