title: Fast live simultaneous multiwavelength four-dimensional optical microscopy.
abstract: Live fluorescence microscopy has the unique capability to probe dynamic processes, linking molecular components and their localization with function. A key goal of microscopy is to increase spatial and temporal resolution while simultaneously permitting identification of multiple specific components. We demonstrate a new microscope platform, OMX, that enables subsecond, multicolor four-dimensional data acquisition and also provides access to subdiffraction structured illumination imaging. Using this platform to image chromosome movement during a complete yeast cell cycle at one 3D image stack per second reveals an unexpected degree of photosensitivity of fluorophore-containing cells. To avoid perturbation of cell division, excitation levels had to be attenuated between 100 and 10,000× below the level normally used for imaging. We show that an image denoising algorithm that exploits redundancy in the image sequence over space and time allows recovery of biological information from the low light level noisy images while maintaining full cell viability with no fading.
authors: Department of Biochemistry and Biophysics, University of California, San Francisco, The Keck Center for Advanced Microscopy, CA 94158-2517, USA.
authors: Peter M
authors: PM
authors: Carlton
authors: Jérôme
authors: J
authors: Boulanger
authors: Charles
authors: C
authors: Kervrann
authors: Jean-Baptiste
authors: JB
authors: Sibarita
authors: Jean
authors: J
authors: Salamero
authors: Susannah
authors: S
authors: Gordon-Messer
authors: Debra
authors: D
authors: Bressan
authors: James E
authors: JE
authors: Haber
authors: Sebastian
authors: S
authors: Haase
authors: Lin
authors: L
authors: Shao
authors: Lukman
authors: L
authors: Winoto
authors: Atsushi
authors: A
authors: Matsuda
authors: Peter
authors: P
authors: Kner
authors: Satoru
authors: S
authors: Uzawa
authors: Mats
authors: M
authors: Gustafsson
authors: Zvi
authors: Z
authors: Kam
authors: David A
authors: DA
authors: Agard
authors: John W
authors: JW
authors: Sedat
mesh_terms: D000465:Algorithms; D000818:Animals; D002470:Cell Survival; D004331:Drosophila melanogaster / Q000166:cytology; D008856:Microscopy, Fluorescence / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000166:cytology; D012984:Software