Abstract
Micropatterned arrays considerably advanced single cell fluorescence time-lapse measurements by providing standardized boundary conditions for thousands of cells in parallel. In these assays, cells are forced to adhere to defined microstructured protein islands separated by passivated, nonadhesive areas. Here we provide a detailed protocol on how to reproducibly fabricate high quality single cell arrays by microscale plasma-initiated protein patterning (μPIPP). Advantages of μPIPP arrays are the ease of preparation and the unrestricted choice of substrates as well as proteins. We demonstrate how the arrays enable the efficient measurement of single cell time trajectories using automated data acquisition and data analysis by example of single cell gene expression after mRNA transfection and time courses of single cell apoptosis. We discuss the more general use of the protocol for assessment of single cell dynamics with the help of fluorescent reporters.
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Acknowledgement
Anita Reiser is supported by a DFG Fellowship through the Graduate School of Quantitative Biosciences Munich (QBM). Support from the European Commission’s 7th Framework Programme through project NanoMILE (Contract No. NMP4-LA-2013-310451) and from the Deutsche Forschungsgemeinschaft via the Nano Initiative Munich (NIM) and SFB 1032 project B01 is gratefully acknowledged.
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Reiser, A., Zorn, M.L., Murschhauser, A., Rädler, J.O. (2018). Single Cell Microarrays Fabricated by Microscale Plasma-Initiated Protein Patterning (μPIPP). In: Ertl, P., Rothbauer, M. (eds) Cell-Based Microarrays. Methods in Molecular Biology, vol 1771. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7792-5_4
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DOI: https://doi.org/10.1007/978-1-4939-7792-5_4
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