Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging

Sandmeyer A, Lachetta M, Sandmeyer H, Hübner W, Huser T, Müller M (2021)
ACS Photonics 8(6): 1639-1648.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Optical nanoscopy is rapidly gaining momentum in the life sciences. Current instruments are, however, often large and expensive, and there is a substantial delay between raw data collection and super-resolved image display. Here, we describe the implementation of a compact, cost-effective, high-speed, structured illumination microscope (SIM), which allows for video-rate super-resolved image reconstructions at imaging rates up to 60 Hz. The instrument is based on a digital micromirror device (DMD) and a global-shutter camera, which enables faster pattern cycles and higher duty cycles than commonly used liquid crystal-based spatial light modulators. In order to utilize a DMD for creating illumination patterns by the coherent superposition of laser beams, we carefully studied its blazed grating effect Through both simulation and experimental determination of system parameters, we identified and optimized its alignment for optimal SIM pattern contrast. Raw image data are collected using inexpensive industry-grade CMOS cameras, while a parallel-computing platform allowed us to reconstruct and visualize living cells in real time. We demonstrate the performance of this system by imaging submicron-sized fluorescent beads diffusing in an aqueous solution, resolving bead-bead interactions in real time. We show that the system is sensitive enough to image intracellular vesicles labeled with fluorescent proteins in fixed cells. We also image dynamic fluctuations of the endoplasmic reticulum (ER), as well as the movement of mitochondria in living osteosarcoma cells, where the cellular organelles are labeled with live cell fluorescent stains.
Stichworte
structured illumination microscopy; digital micromirror devices; super-resolution; live cell imaging; video-rate imaging
Erscheinungsjahr
2021
Zeitschriftentitel
ACS Photonics
Band
8
Ausgabe
6
Seite(n)
1639-1648
eISSN
2330-4022
Page URI
https://pub.uni-bielefeld.de/record/2956276

Zitieren

Sandmeyer A, Lachetta M, Sandmeyer H, Hübner W, Huser T, Müller M. Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging. ACS Photonics. 2021;8(6):1639-1648.
Sandmeyer, A., Lachetta, M., Sandmeyer, H., Hübner, W., Huser, T., & Müller, M. (2021). Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging. ACS Photonics, 8(6), 1639-1648. https://doi.org/10.1021/acsphotonics.0c01937
Sandmeyer, A., Lachetta, M., Sandmeyer, H., Hübner, W., Huser, T., and Müller, M. (2021). Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging. ACS Photonics 8, 1639-1648.
Sandmeyer, A., et al., 2021. Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging. ACS Photonics, 8(6), p 1639-1648.
A. Sandmeyer, et al., “Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging”, ACS Photonics, vol. 8, 2021, pp. 1639-1648.
Sandmeyer, A., Lachetta, M., Sandmeyer, H., Hübner, W., Huser, T., Müller, M.: Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging. ACS Photonics. 8, 1639-1648 (2021).
Sandmeyer, Alice, Lachetta, Mario, Sandmeyer, Hauke, Hübner, Wolfgang, Huser, Thomas, and Müller, Marcel. “Cost-Effective Live Cell Structured Illumination Microscopy with Video-Rate Imaging”. ACS Photonics 8.6 (2021): 1639-1648.

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