Spatial molecular analysis of live cells
Spatially-resolved probing of cells and their microenvironment on biological substrates, is fundamental to understanding cell communication, signaling and growth. We capitalize on the ability of the microfluidic probe to shape and confine biochemicals, and apply micro-scale analytical methods on adherent cells. Our recent work was directed at identifying the amplification status of breast cancer cells in culture, using localized fluorescence in-situ hybridization (FISH) that we term µFISH. The µFISH strategy enables the identification of multiple sub-populations of cells in culture on a given substrate, by multiplexing FISH probes on adjacent regions on a cell monolayer. We have developed another strategy, that locally lyses cells in culture to study their genomic and transcriptomic profiles, a strategy we term Spatialyse. Recently, we studied the gene expression profile of two breast cancer cell lines in co-culture, by lysing cells from the different sub-populations using spatialyse. By analyzing their lysates sequentially, we obtained their unique mRNA signatures. µFISH and Spatialyse can be used sequentially to identify sub-populations of interest and obtain their lysates for molecular analyses of DNA and mRNA.
Kashyap, A., Autebert, J., Delamarche, E. & Kaigala, G. V. Selective local lysis and sampling of live cells for nucleic acid analysis using a microfluidic probe. Sci. Rep. 6, 29579 (2016).
Huber, D., Autebert, J. & Kaigala, G. V. Micro fluorescence in situ hybridization (μFISH) for spatially multiplexed analysis of a cell monolayer. Biomed. Microdevices 18, (2016).