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Single Cell Microarrays Fabricated by Microscale Plasma-Initiated Protein Patterning (μPIPP)

  • Anita Reiser
  • Matthias Lawrence Zorn
  • Alexandra Murschhauser
  • Joachim Oskar Rädler
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1771)

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.

Key words

Microscale plasma-initiated protein patterning (μPIPP) Single-cell analysis High-throughput screening Time-lapse microscopy 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Anita Reiser
    • 1
  • Matthias Lawrence Zorn
    • 1
  • Alexandra Murschhauser
    • 1
  • Joachim Oskar Rädler
    • 1
  1. 1.Center for NanoScience (CeNS), Faculty of PhysicsLudwig-Maximilians-UniversityMunichGermany

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