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Super-Resolution Fluorescence Optical Microscopy: Targeted and Stochastic Read-Out Approaches

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Novel Approaches for Single Molecule Activation and Detection

Part of the book series: Advances in Atom and Single Molecule Machines ((AASMM))

Abstract

This chapter is dedicated to a general overview of some of the emerging and well-established super-resolution techniques recently developed and known as optical nanoscopy and localization precision method. Due to the way of probing the sample, one can consider them as targeted and stochastic-based techniques, respectively. Here, we stress how super-resolution is obtained without violating any physical law, i.e., diffraction. The strong idea behind such approaches, operating in fluorescence contrast mode, is related to the ability of controlling the states, bright/dark or red/blue, of the fluorescent labels being used in order to circumvent the diffraction barrier. Super-resolution is achieved by precluding simultaneous emission of spectrally identical emission of adjacent (<diffraction limit distance) molecules. Also, the evolution of such techniques toward applications on thick (>50 micron thickness) samples is discussed along with correlative microscopy approaches involving scanning probe methods. Examples are given within the neuroscience framework.

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Authors and Affiliations

  1. NanoBioPhotonics—LAMBS, Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genoa, Italy

    Alberto Diaspro, Francesca Cella Zanacchi, Paolo Bianchini & Giuseppe Vicidomini

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  1. Alberto Diaspro
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  2. Francesca Cella Zanacchi
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  3. Paolo Bianchini
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  4. Giuseppe Vicidomini
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Correspondence to Alberto Diaspro .

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Editors and Affiliations

  1. Department of Neuroscience and Neurotechnology, University of Genoa The Italian Institute of Technology, Genoa, Italy

    Fabio Benfenati

  2. Nanostructures Department, The Italian Institute of Technology, Genoa, Italy

    Enzo Di Fabrizio

  3. Neuroscience Area, Scuola Internazionale Superiore di Studi, Trieste, Italy

    Vincent Torre

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Diaspro, A., Zanacchi, F.C., Bianchini, P., Vicidomini, G. (2014). Super-Resolution Fluorescence Optical Microscopy: Targeted and Stochastic Read-Out Approaches. In: Benfenati, F., Di Fabrizio, E., Torre, V. (eds) Novel Approaches for Single Molecule Activation and Detection. Advances in Atom and Single Molecule Machines. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-43367-6_3

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