Video-rate Multi-color Structured Illumination Microscopy with Simultaneous Real-time Reconstruction

Overview

Journal Nat Commun

Specialty Biology

Date 2019 Sep 22

PMID 31541134

Citations 43

Authors

Andreas Markwirth

Mario Lachetta

Viola Monkemoller

Rainer Heintzmann

Wolfgang Hubner

Thomas Huser

Marcel Muller

Affiliations

Soon will be listed here.

Abstract

Super-resolved structured illumination microscopy (SR-SIM) is among the fastest fluorescence microscopy techniques capable of surpassing the optical diffraction limit. Current custom-build instruments are able to deliver two-fold resolution enhancement with high acquisition speed. SR-SIM is usually a two-step process, with raw-data acquisition and subsequent, time-consuming post-processing for image reconstruction. In contrast, wide-field and (multi-spot) confocal techniques produce high-resolution images instantly. Such immediacy is also possible with SR-SIM, by tight integration of a video-rate capable SIM with fast reconstruction software. Here we present instant SR-SIM by VIGOR (Video-rate Immediate GPU-accelerated Open-Source Reconstruction). We demonstrate multi-color SR-SIM at video frame-rates, with less than 250 ms delay between measurement and reconstructed image display. This is achieved by modifying and extending high-speed SR-SIM image acquisition with a new, GPU-enhanced, network-enabled image-reconstruction software. We demonstrate high-speed surveying of biological samples in multiple colors and live imaging of moving mitochondria as an example of intracellular dynamics.

Citing Articles

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