Inaugural Article: Fast live simultaneous multiwavelength four-dimensional optical microscopy
1. Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16016-22. doi:
10.1073/pnas.1004037107. Epub 2010 Aug 12.
Fast live simultaneous multiwavelength four-dimensional optical microscopy.
Carlton PM(1), Boulanger J, Kervrann C, Sibarita JB, Salamero J, Gordon-Messer S,
Bressan D, Haber JE, Haase S, Shao L, Winoto L, Matsuda A, Kner P, Uzawa S,
Gustafsson M, Kam Z, Agard DA, Sedat JW.
(1)Department of Biochemistry and Biophysics, University of California, San
Francisco, The Keck Center for Advanced Microscopy, CA 94158-2517, USA.
Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16005-6.
Nat Methods. 2010 Oct;7(10):782.
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.
PMID: 20705899 [Indexed for MEDLINE]