Journal article
Instant super-resolution imaging in live cells and embryos via analog image processing
Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA1
National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA2
National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA3
National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA4
Existing super-resolution fluorescence microscopes compromise acquisition speed to provide subdiffractive sample information. We report an analog implementation of structured illumination microscopy that enables 3D super-resolution imaging with 145 nm lateral and 350 nm axial resolution, at acquisition speeds up to 100 Hz.
By performing image processing operations optically instead of digitally, we removed the need to capture, store, and combine multiple camera exposures, increasing data acquisition rates 10–100x over other super-resolution microscopes and acquiring and displaying super-resolution images in real-time.
Low excitation intensities allow imaging over hundreds of 2D sections, and combined physical and computational sectioning allow similar depth penetration to confocal microscopy. We demonstrate the capability of our system by imaging fine, rapidly moving structures including motor-driven organelles in human lung fibroblasts and the cytoskeleton of flowing blood cells within developing zebrafish embryos.
Language: | Undetermined |
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Publisher: | Nature Publishing Group US |
Year: | 2014 |
Pages: | 1122-1126 |
Journal subtitle: | Techniques for Life Scientists and Chemists |
ISSN: | 15487105 and 15487091 |
Types: | Journal article |
DOI: | 10.1038/nmeth.2687 |