Fast, long-term, super-resolution imaging with Hessian structured illumination microscopy
Top Cited Papers
- 11 April 2018
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Biotechnology
- Vol. 36 (5), 451-459
- https://doi.org/10.1038/nbt.4115
Abstract
To increase the temporal resolution and maximal imaging time of super-resolution (SR) microscopy, we have developed a deconvolution algorithm for structured illumination microscopy based on Hessian matrixes (Hessian-SIM). It uses the continuity of biological structures in multiple dimensions as a priori knowledge to guide image reconstruction and attains artifact-minimized SR images with less than 10% of the photon dose used by conventional SIM while substantially outperforming current algorithms at low signal intensities. Hessian-SIM enables rapid imaging of moving vesicles or loops in the endoplasmic reticulum without motion artifacts and with a spatiotemporal resolution of 88 nm and 188 Hz. Its high sensitivity allows the use of sub-millisecond excitation pulses followed by dark recovery times to reduce photobleaching of fluorescent proteins, enabling hour-long time-lapse SR imaging of actin filaments in live cells. Finally, we observed the structural dynamics of mitochondrial cristae and structures that, to our knowledge, have not been observed previously, such as enlarged fusion pores during vesicle exocytosis.Keywords
This publication has 45 references indexed in Scilit:
- Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probesProceedings of the National Academy of Sciences of the United States of America, 2012
- Nanoscopy in a Living Mouse BrainScience, 2012
- Calcium Control of Neurotransmitter ReleaseCold Spring Harbor Perspectives in Biology, 2011
- Analysis of Red-Fluorescent Proteins Provides Insight into Dark-State Conversion and PhotodegradationBiophysical Journal, 2011
- Fast live simultaneous multiwavelength four-dimensional optical microscopyProceedings of the National Academy of Sciences of the United States of America, 2010
- A guide to super-resolution fluorescence microscopyThe Journal of cell biology, 2010
- Super-Resolution Fluorescence MicroscopyAnnual Review of Biochemistry, 2009
- Three-Dimensional Resolution Doubling in Wide-Field Fluorescence Microscopy by Structured IlluminationBiophysical Journal, 2008
- Release of small transmitters through kiss-and-run fusion pores in rat pancreatic β cellsCell Metabolism, 2006
- Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolutionProceedings of the National Academy of Sciences of the United States of America, 2005