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

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


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 



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.


  1. 1.
    Jiang X, Bruzewicz DA, Wong AP, Piel M, Whitesides GM (2005) Directing cell migration with asymmetric micropatterns. Proc Natl Acad Sci U S A 102(4):975–978. Scholar
  2. 2.
    Théry M (2010) Micropatterning as a tool to decipher cell morphogenesis and functions. J Cell Sci 123(24):4201–4213. Scholar
  3. 3.
    Piel M, Théry M (2014) Micropatterning in cell biology part a/B/C, vol 119. Academic PressGoogle Scholar
  4. 4.
    Röttgermann PJF, Alberola AP, Radler JO (2014) Cellular self-organization on micro-structured surfaces. Soft Matter 10(14):2397–2404. Scholar
  5. 5.
    Segerer FJ, Röttgermann PJF, Schuster S, Piera Alberola A, Zahler S, Rädler JO (2016) Versatile method to generate multiple types of micropatterns. Biointerphases 11(1):011005. Scholar
  6. 6.
    Segerer FJ, Thüroff F, Piera Alberola A, Frey E, Rädler JO (2015) Emergence and persistence of collective cell migration on small circular micropatterns. Phys Rev Lett 114(22):228102CrossRefGoogle Scholar
  7. 7.
    Röttgermann PFJ, Dawson K, Rädler JO (2016) Time-resolved study of nanoparticle induced apoptosis using microfabricated single cell arrays. Microarrays 5(2):8. Scholar
  8. 8.
    Chatzopoulou EI, Roskopf CC, Sekhavati F, Braciak TA, Fenn NC, Hopfner K-P, Oduncu FS, Fey GH, Rädler JO (2016) Chip-based platform for dynamic analysis of NK cell cytolysis mediated by a triplebody. Analyst 141(7):2284–2295CrossRefGoogle Scholar
  9. 9.
    Ferizi M, Leonhardt C, Meggle C, Aneja MK, Rudolph C, Plank C, Rädler JO (2015) Stability analysis of chemically modified mRNA using micropattern-based single-cell arrays. Lab Chip 15(17):3561–3571. Scholar
  10. 10.
    Picone R, Baum B, McKendry R (2014) Chapter 5 - plasma microcontact patterning (PμCP): a technique for the precise control of surface patterning at small-scale. In: Matthieu P, Manuel T (eds) Methods in cell biology, vol 119. Academic Press, pp 73–90. Scholar
  11. 11.
  12. 12.
    Edelstein A, Amodaj N, Hoover K, Vale R, Stuurman N (2001) Computer control of microscopes using μManager. In: Current protocols in molecular biology. John Wiley & Sons, Inc.
  13. 13.
    Schwarzfischer M, Marr C, Krumsiek J, Hoppe P, Schroeder T, Theis FJ (2011) Efficient fluorescence image normalization for time lapse movies. Proc Microscopic Image Analysis with Applications in Biology 5Google Scholar

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
    Email author
  1. 1.Center for NanoScience (CeNS), Faculty of PhysicsLudwig-Maximilians-UniversityMunichGermany

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