ZFIN ID: ZDB-PUB-131108-27
Instant super-resolution imaging in live cells and embryos via analog image processing
York, A.G., Chandris, P., Nogare, D.D., Head, J., Wawrzusin, P., Fischer, R.S., Chitnis, A., and Shroff, H.
Date: 2013
Source: Nature Methods 10(11): 1122-1126 (Journal)
Registered Authors: Chitnis, Ajay
Keywords: none
MeSH Terms:
  • Animals
  • Embryo, Mammalian/cytology*
  • Microscopy, Fluorescence
PubMed: 24097271 Full text @ Nat. Methods
ABSTRACT

Existing super-resolution fluorescence microscopes compromise acquisition speed to provide subdiffractive sample information. We report an analog implementation of structured illumination microscopy that enables three-dimensional (3D) super-resolution imaging with a lateral resolution of 145 nm and an axial resolution of 350 nm at acquisition speeds up to 100 Hz. By using optical instead of digital image-processing operations, we removed the need to capture, store and combine multiple camera exposures, increasing data acquisition rates 10- to 100-fold 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 spinning-disk 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.

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