Analytical and Bioanalytical Chemistry

, Volume 395, Issue 3, pp 601–609 | Cite as

Plasma stencilling methods for cell patterning

  • Jean-Philippe Frimat
  • Heike Menne
  • Antje Michels
  • Silke Kittel
  • Raffael Kettler
  • Sabine Borgmann
  • Joachim Franzke
  • Jonathan West
Original Paper

Abstract

In this paper we describe plasma stencilling techniques for patterning 10 mammalian cell lines on hydrophobic and cell repellent poly(dimethylsiloxane) (PDMS), methylated glass and bacterial grade polystyrene surfaces. An air plasma produced with a Tesla generator operating at atmospheric pressure was used with microengineered stencils for patterned surface oxidation, selectively transforming the surface to a hydrophilic state to enable cell adhesion and growth. Plasma stencilling obviates the need for directly patterning cell adhesion molecules. Instead, during cell culture, adhesion proteins from the media assemble in a bioactive form on the hydrophilic regions. Critically, the removal of protein patterning prior to cell culture provides the option to also use PDMS–PDMS plasma bonding to incorporate cell patterns within microfluidic systems. Linear patterns were generated using PDMS microchannel stencils, and polyimide stencils with through holes were used for the production of cellular arrays. For the production of smaller cellular arrays, a novel microcapillary-based dielectric barrier discharge system was developed. A numerical method to characterise the cell patterns is also introduced and was used to demonstrate that plasma stencilling is highly effective, with complete patterns confined during long term cell culture (>10 days). In summary, plasma stencilling is simple, rapid, inexpensive, reproducible and a potentially universal cell line patterning capability.

Figure

Microfluidic plasma stencilling for generating cell lines.

Keywords

Cell patterning Plasmas Stencil Poly(dimethylsiloxane) Dielectric barrier discharge Microfluidics 

Supplementary material

216_2009_2824_Fig7_ESM.gif (15 kb)
Fig. S1

White light interferometry analysis of polystyrene (BGPS) surface roughening caused by plasma stencilling for 1 s (a), 3 s (b) and 5 s (c) (GIF 14.6 kb)

216_2009_2824_Fig7_ESM.tif (7.6 mb)
Fig. S1High resolution image file (TIFF 7.58 mb).
216_2009_2824_Fig8_ESM.gif (48 kb)
Fig. S2

SEM image of BGPS after prolonged (5 s) plasma treatment (GIF 47.6 kb)

216_2009_2824_Fig8_ESM.tif (2.3 mb)
Fig. S2High resolution image file (TIFF 2.30 mb).
216_2009_2824_Fig9_ESM.gif (69 kb)
Fig. S3

PDMS microfluidic system incorporating patterned cells. Plasma stencilling was used for both cell patterning and to enable bonding between both PDMS layers (GIF 69 kb)

216_2009_2824_Fig9_ESM.tif (6.6 mb)
Fig. S3High resolution image file (TIFF 6.57 mb).
216_2009_2824_MOESM1_ESM.docx (12 kb)
Table S1 (DOC 12 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Jean-Philippe Frimat
    • 1
  • Heike Menne
    • 1
  • Antje Michels
    • 1
  • Silke Kittel
    • 1
  • Raffael Kettler
    • 1
  • Sabine Borgmann
    • 1
  • Joachim Franzke
    • 1
  • Jonathan West
    • 1
  1. 1.ISAS—Institute for Analytical SciencesDortmundGermany

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