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Microfluidic Platform for Parallel Single Cell Analysis for Diagnostic Applications

  • Séverine Le GacEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1547)

Abstract

Cell populations are heterogeneous: they can comprise different cell types or even cells at different stages of the cell cycle and/or of biological processes. Furthermore, molecular processes taking place in cells are stochastic in nature. Therefore, cellular analysis must be brought down to the single cell level to get useful insight into biological processes, and to access essential molecular information that would be lost when using a cell population analysis approach. Furthermore, to fully characterize a cell population, ideally, information both at the single cell level and on the whole cell population is required, which calls for analyzing each individual cell in a population in a parallel manner. This single cell level analysis approach is particularly important for diagnostic applications to unravel molecular perturbations at the onset of a disease, to identify biomarkers, and for personalized medicine, not only because of the heterogeneity of the cell sample, but also due to the availability of a reduced amount of cells, or even unique cells. This chapter presents a versatile platform meant for the parallel analysis of individual cells, with a particular focus on diagnostic applications and the analysis of cancer cells. We first describe one essential step of this parallel single cell analysis protocol, which is the trapping of individual cells in dedicated structures. Following this, we report different steps of a whole analytical process, including on-chip cell staining and imaging, cell membrane permeabilization and/or lysis using either chemical or physical means, and retrieval of the cell molecular content in dedicated channels for further analysis. This series of experiments illustrates the versatility of the herein-presented platform and its suitability for various analysis schemes and different analytical purposes.

Key words

Single cell Parallelization Microfluidics Circulating tumor cells Cell trapping Cell permeabilization 

Notes

Acknowledgment

The work presented in this chapter has been conducted in part by Floris van den Brink and Elmar Gool. Johan Bomer, Daniël Wijnperlé, and Hans de Boer are thanked for technical assistance (mold fabrication and development of the setup). Prof. Edwin Carlen is acknowledged for his supervision in the first year of the project. This work was supported by NanoNext NL, a micro and nanotechnology consortium of the Government of the Netherlands and 130 partners as well as by the company OGT (Oxford Gene Technology).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Applied Microfluidics for BioEngineering ResearchMESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Engineering and Technical Medicine, University of TwenteEnschedeThe Netherlands

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