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