High speed multi-plane super-resolution structured illumination microscopy of living cells using an image-splitting prism

Descloux, A.; Müller, M.; Navikas, V.; Markwirth, A.; Van den Eynde, R.; Lukes, T.; Hübner, W.; Lasser, T.; Radenovic, A.; Dedecker, P.; Huser, T.

doi:10.1101/773440

bioRxiv · 10.1101/773440

High speed multi-plane super-resolution structured illumination microscopy of living cells using an image-splitting prism

Descloux, A.·Müller, M.·Navikas, V.·Markwirth, A.·Van den Eynde, R.·Lukes, T.·Hübner, W.·Lasser, T.·Radenovic, A.·Dedecker, P.·Huser, T.

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Abstract

Super-resolution structured illumination microscopy (SR-SIM) can be conducted at video-rate acquisition speeds when combined with high-speed spatial light modulators and sCMOS cameras, rendering it particularly suitable for live cell imaging. If, however, three-dimensional (3D) information is desired, the sequential acquisition of vertical image stacks employed by current setups significantly slows down the acquisition process. In this work we present a multi-plane approach to SR-SIM that overcomes this slowdown via the simultaneous acquisition of multiple object planes, employing a recently introduced multi-plane image splitting prism combined with high-speed SR-SIM illumination. This strategy requires only the introduction of a single optical element and the addition of a second camera to acquire a laterally super-resolved three-dimensional image stack. We demonstrate the performance of multi-plane SR-SIM by applying this instrument to the dynamics of live mitochondrial network.

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Descloux, A., Müller, M., Navikas, V., Markwirth, A., Van den Eynde, R., Lukes, T., Hübner, W., Lasser, T., Radenovic, A., Dedecker, P., Huser, T.. 2019-09-18. High speed multi-plane super-resolution structured illumination microscopy of living cells using an image-splitting prism. https://doi.org/10.1101/773440

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