Hydrodynamic guiding for addressing subsets of immobilized cells and molecules in microfluidic systems.
BACKGROUND
The interest in microfluidics and surface patterning is increasing as the use of these technologies in diverse biomedical applications is substantiated. Controlled molecular and cellular surface patterning is a costly and time-consuming process. Methods for keeping multiple separate experimental conditions on a patterned area are, therefore, needed to amplify the amount of biological information that can be retrieved from a patterned surface area. We describe, in three examples of biomedical applications, how this can be achieved in an open microfluidic system, by hydrodynamically guiding sample fluid over biological molecules and living cells immobilized on a surface.


RESULTS
A microfluidic format of a standard assay for cell-membrane integrity showed a fast and dose-dependent toxicity of saponin on mammalian cells. A model of the interactions of human mononuclear leukocytes and endothelial cells was established. By contrast to static adhesion assays, cell-cell adhesion in this dynamic model depended on cytokine-mediated activation of both endothelial and blood cells. The microfluidic system allowed the use of unprocessed blood as sample material, and a specific and fast immunoassay for measuring the concentration of C-reactive protein in whole blood was demonstrated.


CONCLUSIONS
The use of hydrodynamic guiding made multiple and dynamic experimental conditions on a small surface area possible. The ability to change the direction of flow and produce two-dimensional grids can increase the number of reactions per surface area even further. The described microfluidic system is widely applicable, and can take advantage of surfaces produced by current and future techniques for patterning in the micro- and nanometer scale.
Brevig
Thomas
T
Department of Molecular Characterization, Biotechnological Institute, Kogle Allé 2, DK-2970 Hørsholm, Denmark. tbr@bioteknologisk.dk
Krühne
Ulrich
U
Kahn
Rachel A
RA
Ahl
Thomas
T
Beyer
Michael
M
Pedersen
Lars H
LH
D000818:Animals; D002097:C-Reactive Protein / Q000378:metabolism; D016466:CHO Cells; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008832:Microchemistry / Q000379:methods*; D012503:Saponins / Q000494:pharmacology; D013499:Surface Properties; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018925:Chemokines; D012503:Saponins; D014409:Tumor Necrosis Factor-alpha; D002097:C-Reactive Protein