Fast live simultaneous multiwavelength four-dimensional optical microscopy

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...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2010-09, Vol.107 (37), p.16016-16022
Main Authors: Carlton, Peter M., Boulanger, Jérôme, Kervrann, Charles, Sibarita, Jean-Baptiste, Salamero, Jean, Gordon-Messer, Susannah, Bressan, Debra, Haber, James E., Haase, Sebastian, Shao, Lin, Winoto, Lukman, Matsuda, Atsushi, Kner, Peter, Uzawa, Satoru, Gustafsson, Mats, Kam, Zvi, Agard, David A., Sedat, John W.
Format: Article
Language:English
Subjects:
Algorithms
Animals
Biological Sciences
Cell cycle
Cell division
Cell Survival
Cellular Biology
Chromosomes
Computer Science
Drosophila melanogaster - cytology
Fluorescence
Image Processing
Imaging
Life Sciences
Luminous intensity
Microscopy
Microscopy, Fluorescence - methods
Optics
Phototoxicity
Physics
Saccharomyces cerevisiae - cytology
Signal noise
Software
Viability
Yeast
Yeasts
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 sub-diffraction 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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1004037107