title: Real-time monitoring of cell migration, phagocytosis and cell surface receptor dynamics using a novel, live-cell opto-microfluidic technique.
abstract: We report an opto-microfluidic method for continuous and non-interfering monitoring of cell movement and dynamic molecular processes in living cells enabled by the microfluidic "Lab-in-a-Trench" (LiaT) platform. To demonstrate real-time monitoring of heterogeneous cell-cell interactions, cell tracking and agent-induced cell activation dynamics, we observe phagocytosis of Escherichia coli by murine macrophages, migration of active macrophages and LPS-induced CD86 expression in macrophages. The visualization of phagocytosis is facilitated through the loading of green fluorescent protein (GFP) expressing E. coli to the array of cell capture modules before the introduction of macrophages. Simple migration tracking of active macrophages is enabled by a spatio-temporal control of the environment conditions within the LiaT platform. Furthermore, we report an interference-free monitoring of non-modified, endogenous changes in protein expression on the surface of living cells using traditional, antibody immuno-reagents. Throughout the experiment, murine macrophages were captured in the LiaT device and exposed to sub-background levels of fluorescently labeled anti-CD86 antibody. Upon lipopolysaccharide (LPS) stimulation, CD86 changes were visualized in real-time by time-lapse microscopy. This novel opto-microfluidic effect is controlled by the equilibrium of convective-diffusive replenishment of fluorescently labeled antibodies and antibody affinity. Overall, our non-interfering analysis method allows the studying of active cellular processes and endogenous protein dynamics in live cells in a simple and cost-efficient manner.
authors.affiliation: Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland. Electronic address: gregor.kijanka@dcu.ie.
authors.forename: Gregor S
authors.initials: GS
authors.lastname: Kijanka
authors.affiliation: Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Ivan K
authors.initials: IK
authors.lastname: Dimov
authors.affiliation: Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland; School of Physical Sciences, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Robert
authors.initials: R
authors.lastname: Burger
authors.affiliation: Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland; School of Physical Sciences, National Centre for Sensor Research, Dublin City University, Glasnevin, Dublin 9, Ireland. Electronic address: jens.ducree@dcu.ie.
authors.forename: Jens
authors.initials: J
authors.lastname: Ducrée
mesh_terms: D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D010587:Phagocytosis; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013997:Time Factors
chemical_list: D011956:Receptors, Cell Surface
keywords: Cell migration; Cell surface marker expression; Microfluidics; Phagocytosis; Real-time monitoring; Time-lapse microscopy