A system and methodology for high-content visual screening of individual intact living cells in suspension

Olivier Renaud; Rainer Heintzmann; Asier Sáez-Cirión; Thomas Schnelle; Torsten Mueller; Spencer Shorte

doi:10.1117/12.699000

19 February 2007 A system and methodology for high-content visual screening of individual intact living cells in suspension

Olivier Renaud, Rainer Heintzmann, Asier Sáez-Cirión, Thomas Schnelle, Torsten Mueller, Spencer Shorte

Author Affiliations +

Proceedings Volume 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V; 64410Q (2007) https://doi.org/10.1117/12.699000
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

Citation Download Citation

Olivier Renaud, Rainer Heintzmann, Asier Sáez-Cirión, Thomas Schnelle, Torsten Mueller, and Spencer Shorte "A system and methodology for high-content visual screening of individual intact living cells in suspension", Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64410Q (19 February 2007); https://doi.org/10.1117/12.699000

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