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Micropatterned "adherent/repellent" glass surfaces for studying the spreading kinetics of individual red blood cells onto protein-decorated substrates

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Abstract

We report in this paper two simple and effective methods to decorate glass surfaces that enable protein micropatterning and subsequent spatially controlled adhesion of cells. The first method combines simultaneously the potentialities of two existing techniques, namely microcontact printing (μCP) and microfluidic networks (μFN) to achieve dual protein patterning in a single step. The second method is mainly based on the well-known property of poly(ethylene glycol) (PEG) to resist against protein adsorption. Both approaches were used to produce heterogeneous surfaces on which micron-size or submicronic streptavidin-coated lines alternate with cell-repellent areas. We first describe the implementation of the two methods and discuss the main pitfalls to avoid. Then, using these templates, we have monitored the kinetics of attachment of individual biotinylated (i.e. "attractant" towards streptavidin) red blood cells by directly measuring the propagation velocity of the adhesion front. Depending on the surface density of biotin, we found two distinct regimes, in agreement with existing theoretical models.

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Abbreviations

μCP:

microcontact printing

μFN:

microfluidic networks

BSA:

bovine serum albumin

CB:

carbonate-bicarbonate buffer

EDA:

N-[3-(trimethoxysilyl)propyl]ethylenediamine

NHS:

N-hydroxysuccinimidyl

PBS:

phosphate buffered saline

PDMS:

poly(dimethylsiloxane)

PEG:

poly(ethylene glycol)

RICM:

reflection interference contrast microscopy

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Acknowledgements

We thank Axel Buguin for his advice on lithography, Françoise Brochard-Wyart for stimulating discussions, and Jacques Prost for supporting our ongoing project. The work was supported by the CNRS through the ACI "Physicochimie de la Matière Complexe 2000" and "Nano-Objet Individuel 2001" programs as well as by a grant from the La Ligue Nationale contre le Cancer (N/ref. 75/01-RS/83). One of us (O.R.) thanks the ARC for the one-year fellowship (no. Ml/MLD/CM-P01/5). We also received generous help from the Institut Curie.

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Correspondence to Pierre Nassoy.

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Cuvelier, D., Rossier, O., Bassereau, P. et al. Micropatterned "adherent/repellent" glass surfaces for studying the spreading kinetics of individual red blood cells onto protein-decorated substrates. Eur Biophys J 32, 342–354 (2003). https://doi.org/10.1007/s00249-003-0282-2

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