Superresolution upgrade for confocal spinning disk systems using image scanning microscopy (Conference Presentation)

Sebastian Isbaner; Dirk Hähnel; Ingo Gregor; Jörg Enderlein

doi:10.1117/12.2255813

24 April 2017 Superresolution upgrade for confocal spinning disk systems using image scanning microscopy (Conference Presentation)

Sebastian Isbaner, Dirk Hähnel, Ingo Gregor, Jörg Enderlein

Author Affiliations +

Proceedings Volume 10071, Single Molecule Spectroscopy and Superresolution Imaging X; 100710F (2017) https://doi.org/10.1117/12.2255813
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

Confocal Spinning Disk Systems are widely used for 3D cell imaging because they offer the advantage of optical sectioning at high framerates and are easy to use. However, as in confocal microscopy, the imaging resolution is diffraction limited, which can be theoretically improved by a factor of 2 using the principle of Image Scanning Microscopy (ISM) [1]. ISM with a Confocal Spinning Disk setup (CSDISM) has been shown to improve contrast as well as lateral resolution (FWHM) from 201 ± 20 nm to 130 ± 10 nm at 488 nm excitation. A minimum total acquisition time of one second per ISM image makes this method highly suitable for 3D live cell imaging [2]. Here, we present a multicolor implementation of CSDISM for the popular Micro-Manager Open Source Microscopy platform. Since changes in the optical path are not necessary, this will allow any researcher to easily upgrade their standard Confocal Spinning Disk system at remarkable low cost (~5000 USD) with an ISM superresolution option. [1]. Müller, C.B. and Enderlein, J. Image Scanning Microscopy. Physical Review Letters 104, (2010). [2]. Schulz, O. et al. Resolution doubling in fluorescence microscopy with confocal spinning-disk image scanning microscopy. Proceedings of the National Academy of Sciences of the United States of America 110, 21000-5 (2013).

Conference Presentation

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Sebastian Isbaner, Dirk Hähnel, Ingo Gregor, and Jörg Enderlein "Superresolution upgrade for confocal spinning disk systems using image scanning microscopy (Conference Presentation)", Proc. SPIE 10071, Single Molecule Spectroscopy and Superresolution Imaging X, 100710F (24 April 2017); https://doi.org/10.1117/12.2255813

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KEYWORDS

Confocal microscopy

Microscopy

Super resolution

3D acquisition

3D image processing

Image resolution

Imaging systems

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