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Optics Express

Optics Express

  • Vol. 17, Iss. 7 — Mar. 30, 2009
  • pp: 5585–5594

Automated focusing of nuclei for time lapse experiments on single cells using holographic optical tweezers

Emma Eriksson, David Engström, Jan Scrimgeour, and Mattias Goksör  »View Author Affiliations


Optics Express, Vol. 17, Issue 7, pp. 5585-5594 (2009)
http://dx.doi.org/10.1364/OE.17.005585


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Abstract

Experiments on single cells are currently gaining more and more interest. Single cell studies often concerns the spatio-temporal distribution of fluorescent proteins inside living cells, visualized using fluorescence microscopy. In order to extract quantitative information from such experiments it is necessary to image the sample with high spatial and temporal resolution while keeping the photobleaching to a minimum. The analysis of the spatial distribution of proteins often requires stacks of images at each time point, which exposes the sample to unnecessary amounts of excitation light. In this paper we show how holographic optical tweezers combined with image analysis can be used to optimize the axial position of trapped cells in an array in order to bring the nuclei into a single imaging plane, thus eliminating the need for stacks of images and consequently reducing photobleaching. This allows more images to be collected, as well as increasing the time span and/or the time resolution in time lapse studies of single cells.

© 2009 Optical Society of America

OCIS Codes
(090.2890) Holography : Holographic optical elements
(170.1530) Medical optics and biotechnology : Cell analysis
(170.4520) Medical optics and biotechnology : Optical confinement and manipulation
(180.2520) Microscopy : Fluorescence microscopy
(230.6120) Optical devices : Spatial light modulators
(350.4855) Other areas of optics : Optical tweezers or optical manipulation

ToC Category:
Optical Trapping and Manipulation

History
Original Manuscript: February 27, 2009
Revised Manuscript: March 18, 2009
Manuscript Accepted: March 20, 2009
Published: March 24, 2009

Virtual Issues
Vol. 4, Iss. 5 Virtual Journal for Biomedical Optics

Citation
Emma O. Eriksson, David Engström, Jan Scrimgeour, and Mattias Goksör, "Automated focusing of nuclei for time lapse experiments on single cells using holographic optical tweezers," Opt. Express 17, 5585-5594 (2009)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-17-7-5585


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