Multiview microscopy of single cells through microstructure-based indirect optical manipulation

Vizsnyiczai, G., Búzás, A., Aekbote, B. L. , Fekete, T., Grexa, I., Ormos, P. and Kelemen, L. (2020) Multiview microscopy of single cells through microstructure-based indirect optical manipulation. Biomedical Optics Express, 11(2), pp. 945-962. (doi: 10.1364/BOE.379233)

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Fluorescent observation of cells generally suffers from the limited axial resolution due to the elongated point spread function of the microscope optics. Consequently, three-dimensional imaging results in axial resolution that is several times worse than the transversal. The optical solutions to this problem usually require complicated optics and extreme spatial stability. A straightforward way to eliminate anisotropic resolution is to fuse images recorded from multiple viewing directions achieved mostly by the mechanical rotation of the entire sample. In the presented approach, multiview imaging of single cells is implemented by rotating them around an axis perpendicular to the optical axis by means of holographic optical tweezers. For this, the cells are indirectly trapped and manipulated with special microtools made with two-photon polymerization. The cell is firmly attached to the microtool and is precisely manipulated with 6 degrees of freedom. The total control over the cells' position allows for its multiview fluorescence imaging from arbitrarily selected directions. The image stacks obtained this way are combined into one 3D image array with a multiview image processing pipeline resulting in isotropic optical resolution that approaches the lateral diffraction limit. The presented tool and manipulation scheme can be readily applied in various microscope platforms.

Item Type:Articles
Additional Information:Funding: Ministry of Finance (GINOP-2.3.2-15-2016-00001, GINOP-2.3.3-15-2016-00040); Horizon 2020 Framework Programme (654148 Laserlab-Europe); Nvidia (GPU grant program).
Glasgow Author(s) Enlighten ID:Aekbote, Dr Badri
Authors: Vizsnyiczai, G., Búzás, A., Aekbote, B. L., Fekete, T., Grexa, I., Ormos, P., and Kelemen, L.
College/School:College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Biomedical Optics Express
Publisher:Optical Society of America
ISSN (Online):2156-7085
Published Online:16 January 2020
Copyright Holders:Copyright © 2020 The Optical Society
First Published:First published in Biomedical Optics Express 11(2): 945-962
Publisher Policy:Reproduced in accordance with the publisher copyright policy
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