Open AccessAutomated focusing of nuclei for time lapse experiments on single cells using holographic optical tweezers
Author(s) -
Emma Eriksson,
David Freeman Engstrom,
Jan Scrimgeour,
Mattias Goksör
Publication year - 2009
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.17.005585
Subject(s) - photobleaching , optical tweezers , holography , optics , live cell imaging , microscopy , image resolution , temporal resolution , light sheet fluorescence microscopy , resolution (logic) , cardinal point , fluorescence microscope , materials science , physics , fluorescence , computer science , chemistry , artificial intelligence , biochemistry , cell
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.