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Proceedings Paper

A system and methodology for high-content visual screening of individual intact living cells in suspension
Author(s): Olivier Renaud; Rainer Heintzmann; Asier Sáez-Cirión; Thomas Schnelle; Torsten Mueller; Spencer Shorte
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Paper Abstract

Three dimensional imaging provides high-content information from living intact biology, and can serve as a visual screening cue. In the case of single cell imaging the current state of the art uses so-called "axial through-stacking". However, three-dimensional axial through-stacking requires that the object (i.e. a living cell) be adherently stabilized on an optically transparent surface, usually glass; evidently precluding use of cells in suspension. Aiming to overcome this limitation we present here the utility of dielectric field trapping of single cells in three-dimensional electrode cages. Our approach allows gentle and precise spatial orientation and vectored rotation of living, non-adherent cells in fluid suspension. Using various modes of widefield, and confocal microscope imaging we show how so-called "microrotation" can provide a unique and powerful method for multiple point-of-view (three-dimensional) interrogation of intact living biological micro-objects (e.g. single-cells, cell aggregates, and embryos). Further, we show how visual screening by micro-rotation imaging can be combined with micro-fluidic sorting, allowing selection of rare phenotype targets from small populations of cells in suspension, and subsequent one-step single cell cloning (with high-viability). Our methodology combining high-content 3D visual screening with one-step single cell cloning, will impact diverse paradigms, for example cytological and cytogenetic analysis on haematopoietic stem cells, blood cells including lymphocytes, and cancer cells.

Paper Details

Date Published: 19 February 2007
PDF: 10 pages
Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64410Q (19 February 2007); doi: 10.1117/12.699000
Show Author Affiliations
Olivier Renaud, Institut Pasteur (France)
Rainer Heintzmann, King's College London (United Kingdom)
Asier Sáez-Cirión, Institut Pasteur (France)
Thomas Schnelle, Evotec Technologies GmbH (Germany)
Torsten Mueller, Evotec Technologies GmbH (Germany)
Spencer Shorte, Institut Pasteur (France)

Published in SPIE Proceedings Vol. 6441:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
Daniel L. Farkas; Robert C. Leif; Dan V. Nicolau, Editor(s)

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