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A High‐throughput Automated Microfluidic Platform for Live Cell Rolling and Dynamic Adhesion Assays
Author(s) -
Conant Carolyn Grace,
Schwartz Michael,
IonescuZanetti Cristian,
Jensen Jeff
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.448
Subject(s) - adhesion , microfluidics , throughput , cell adhesion , shear stress , laminar flow , chemistry , nanotechnology , biophysics , materials science , computer science , biology , composite material , mechanics , physics , telecommunications , wireless
A large number of processes underlying immune system biology are dependent on leukocyte adhesion and neutrophil rolling phenomena. Laminar flow chambers are the standard tool in vitro for studying cellular adhesion under shear stress, but are of limited use for assays with high numbers of variables, eg./ chemical library screens, because of inherent low throughput. We present development of a disposable microfluidic platform which improves upon throughput, experimental control, and setup time for cell adhesion and cell rolling assays. Rolling assays were performed in the presence or absence of divalent cations on this platform on either VCAM‐1 or PNAd. Consistent with established data (Chen, et al, 2004), no adhesion or rolling was observed in the absence of divalent ions. Buffer conditions with magnesium (Mg) resulted in cell adherence with no rolling; addition of calcium (Ca) with or without Mg resulted in slow rolling. Jurkat cell adhesion strength to fibronection under shear flow and adhesion dose response were also successfully assessed using this platform. Principal advantages demonstrated with the BioFlux 200™ system were assay parallelization, real‐time imaging, minimized reagent consumption, precise shear control, integrated image acquisition, and automated cell tracking analysis. Future modifications to the system will include expansion to a 96‐well plate footprint.