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Cell analysis using a multiple internal reflection photonic lab-on-a-chip

Nature Protocols volume 6, pages 1642–1655 (2011)Cite this article

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Abstract

Here we present a protocol for analyzing cell cultures using a photonic lab-on-a-chip (PhLoC). By using a broadband light source and a spectrometer, the spectrum of a given cell culture with an arbitrary population is acquired. The PhLoC can work in three different regimes: light scattering (using label-free cells), light scattering plus absorption (using stained cells) and, by subtraction of the two former regimes, absorption (without the scattering band). The acquisition time of the PhLoC is ∼30 ms. Hence, it can be used for rapid cell counting, dead/live ratio estimation or multiparametric measurements through the use of different dyes. The PhLoC, including microlenses, micromirrors and microfluidics, is simply fabricated in a single-mask process (by soft lithographic methods) using low-cost materials. Because of its low cost it can easily be implemented for point-of-care applications. From raw substrates to final results, this protocol can be completed in 29 h.

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Figure 1: Schematic representation of the merging of micro-optics and microfluidics.
Figure 2: Layout of the mask used for implementing the PhLoC system.
Figure 3: Fabrication steps.
Figure 4: Picture of the completed PhLoC, having self-alignment microchannels for accurate fiber optics positioning, microlenses and air mirrors.
Figure 5: Characterization in the LS regime.
Figure 6: Characterization in the LS + ABS regime.
Figure 7: Characterization in the ABS regime obtained by subtraction of the spectra obtained in LS and LS + ABS regimes.
Figure 8: Measurement of the monocytes dead/live ratio.

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Acknowledgements

The research leading to these results has received funding from the European Research Council under the European Community's Seventh Framework Programme (FP7/2007-2013)/ERC grant agreement no. 209243 and also from the IST Programme (P. CEZANNE, IST-2-IP-031867). We thank the research group FOR 856, mikroPART 'Mikrosysteme für partikuläre Life-Science-Produkte', for support of this work. Discussions with A. Voigt and G. Gruetzner of Microresist (Germany) regarding the fabrication steps are highly appreciated.

AUTHOR CONTRIBUTIONS

J.V.-P., E.F.-R. and B.I. conducted the experiments. E.F.-R. and C.N. prepared the cell culture. S.D., S.B. and A.L. designed and fabricated the PhLoC. J.A.P., C.D., S.B. and A.L. defined the broadband screening protocol. A.L. supervised the project at all stages.

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Authors and Affiliations

  1. Centre Nacional de Microelectrònica (IMB-CNM, CSIC), Barcelona, Spain

    Jordi Vila-Planas, Bergoi Ibarlucea, Jose A Plaza, Carlos Domínguez & Andreu Llobera

  2. Departament de Biologia Cellular, Fisiologia i Immunologia, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain

    Elisabet Fernández-Rosas & Carme Nogués

  3. Institut für Mikrotechnik, Technische Universität Braunschweig, Braunschweig, Germany

    Stefanie Demming, Stephanus Büttgenbach & Andreu Llobera

Authors
  1. Jordi Vila-Planas
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  2. Elisabet Fernández-Rosas
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  9. Andreu Llobera
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Correspondence to Andreu Llobera.

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Vila-Planas, J., Fernández-Rosas, E., Ibarlucea, B. et al. Cell analysis using a multiple internal reflection photonic lab-on-a-chip. Nat Protoc 6, 1642–1655 (2011). https://doi.org/10.1038/nprot.2011.383

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