Implementation of a 4Pi-SMS super-resolution microscope

Jingyu Wang, Edward S. Allgeyer, George Sirinakis, Yongdeng Zhang, Kevin Hu, Mark D. Lessard, Yiming Li, Robin Diekmann, Michael A. Phillips, Ian M. Dobbie, Jonas Ries, Martin J. Booth, Joerg Bewersdorf

Published: 2020-12-15 DOI: 10.1038/s41596-020-00428-7

4Pi-SMS microscopy

super-resolution

fluorescence microscopy

optomechanical assembly

troubleshooting guide

AI 解读

Genetic manipulation and immortalized culture of ex vivo primary human germinal center B cells

The cell-line-derived subcutaneous tumor model in preclinical cancer research

Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous pre- and postsynaptic recording at cortical synapses

NovoSpaRc: flexible spatial reconstruction of single-cell gene expression with optimal transport

Genome-wide pooled CRISPR screening in neurospheres

High-throughput cultivation and identification of bacteria from the plant root microbiota

Viral crosslinking and solid-phase purification enables discovery of ribonucleoprotein complexes on incoming RNA virus genomes

Mitochondrial single-cell ATAC-seq for high-throughput multi-omic detection of mitochondrial genotypes and chromatin accessibility

Addendum: Micro-CT acquisition and image processing to track and characterize pulmonary nodules in mice

Multiplexed single-cell proteomics using SCoPE2

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The development of single-molecule switching (SMS) fluorescence microscopy (also called single-molecule localization microscopy) over the last decade has enabled researchers to image cell biological structures at unprecedented resolution. Using two opposi
Introduction
Materials
Procedure
Troubleshooting
Timing
Anticipated results
References
ACKNOWLEDGMENTS

Shtengel, G. et al. Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure. Proc. Natl Acad. Sci. USA 106, 3125–3130 (2009).
Aquino, D. et al. Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores. Nat. Methods 8, 353–359 (2011).
Wang, G., Hauver, J., Thomas, Z., Darst, S. A. & Pertsinidis, A. Single-molecule real-time 3D imaging of the transcription cycle by modulation interferometry. Cell 167, 1839–1852.e21 (2016).
Huang, F. et al. Ultra-high resolution 3D imaging of whole cells. Cell 166, 1028–1040 (2016).
Del Viso, F. et al. Congenital heart disease genetics uncovers context-dependent organization and function of nucleoporins at cilia. Dev. Cell 38, 478–492 (2016).
Zhang, Y., Lara-Tejero, M., Bewersdorf, J. & Galan, J. E. Visualization and characterization of individual type III protein secretion machines in live bacteria. Proc. Natl Acad. Sci. USA 114, 6098–6103 (2017).
Hwang, J. Y. et al. Dual sensing of physiologic pH and calcium by EFCAB9 regulates sperm motility. Cell 177, 1480–1494.e19 (2019).
Schroeder, L. K. et al. Dynamic nanoscale morphology of the ER surveyed by STED microscopy. J. Cell Biol 218, 83–96 (2019).
Zhang, Y. et al. Nanoscale subcellular architecture revealed by multicolor three-dimensional salvaged fluorescence imaging. Nat. Methods 17, 225–231 (2020).
Karanastasis, A. A. et al. 3D mapping of nanoscale crosslink heterogeneities in microgels. Mater. Horiz. 5, 1130–1136 (2018).
Brown, T. A. et al. Superresolution fluorescence imaging of mitochondrial nucleoids reveals their spatial range, limits, and membrane interaction. Mol. Cell. Biol. 31, 4994–5010 (2011).
Schermelleh, L. et al. Super-resolution microscopy demystified. Nat. Cell Biol. 21, 72–84 (2019).
Barentine, A. E. S. et al. 3D multicolor nanoscopy at 10,000 cells a day. Preprint at https://www.biorxiv.org/content/10.1101/606954v1 (2019).
Booth, M. J. Adaptive optical microscopy: the ongoing quest for a perfect image. Light Sci. Appl. 3, e165 (2014).
Burke, D., Patton, B., Huang, F., Bewersdorf, J. & Booth, M. J. Adaptive optics correction of specimen-induced aberrations in single-molecule switching microscopy. Optica 2, 177–185 (2015).
Van Engelenburg, S. B. et al. Distribution of ESCRT machinery at HIV assembly sites reveals virus scaffolding of ESCRT subunits. Science 343, 653–656 (2014).
Buttler, C. A. et al. Single molecule fate of HIV-1 envelope reveals late-stage viral lattice incorporation. Nat. Commun. 9, 1861 (2018).
Liu, S. & Huang, F. Enhanced 4Pi single-molecule localization microscopy with coherent pupil based localization. Commun. Biol. 3, 220 (2020).
Li, Y. et al. Accurate 4Pi single-molecule localization using an experimental PSF model. Opt. Lett. 45, 3765–3768 (2020).
Li, X. et al. Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM. Nat. Methods 10, 584–590 (2013).
Wang, Y. et al. Localization events-based sample drift correction for localization microscopy with redundant cross-correlation algorithm. Opt. Express 22, 15982–15991 (2014).
Li, Y. et al. Real-time 3D single-molecule localization using experimental point spread functions. Nat. Methods 15, 367–369 (2018).
Thevathasan, J. V. et al. Nuclear pores as versatile reference standards for quantitative superresolution microscopy. Nat. Methods 16, 1045–1053 (2019).
Antonello, J., Wang, J., He, C., Phillips, M. & Booth, M. Interferometric calibration of a deformable mirror. Zenodo. 19 March 2020 (accessed 21 November 2020) https://zenodo.org/record/3714951#.X6RM2lNKgdU

Implementation of a 4Pi-SMS super-resolution microscope

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