title: Analysing calcium signalling of cells under high shear flows using discontinuous dielectrophoresis.
abstract: Immobilisation of cells is an important feature of many cellular assays, as it enables the physical/chemical stimulation of cells; whilst, monitoring cellular processes using microscopic techniques. Current approaches for immobilising cells, however, are hampered by time-consuming processes, the need for specific antibodies or coatings, and adverse effects on cell integrity. Here, we present a dielectrophoresis-based approach for the robust immobilisation of cells, and analysis of their responses under high shear flows. This approach is quick and label-free, and more importantly, minimises the adverse effects of electric field on the cell integrity, by activating the field for a short duration of 120 s, just long enough to immobilise the cells, after which cell culture media (such as HEPES) is flushed through the platform. In optimal conditions, at least 90% of the cells remained stably immobilised, when exposed to a shear stress of 63 dyn/cm(2). This approach was used to examine the shear-induced calcium signalling of HEK-293 cells expressing a mechanosensitive ion channel, transient receptor potential vaniloid type 4 (TRPV4), when exposed to the full physiological range of shear stress.
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, Australia.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Soffe
authors.affiliation: Health Innovations Research Institute and School of Medical Sciences, RMIT University, Melbourne, Australia.
authors.forename: Sara
authors.initials: S
authors.lastname: Baratchi
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, Australia.
authors.forename: Shi-Yang
authors.initials: SY
authors.lastname: Tang
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, Australia.
authors.forename: Mahyar
authors.initials: M
authors.lastname: Nasabi
authors.affiliation: Health Innovations Research Institute and School of Medical Sciences, RMIT University, Melbourne, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: McIntyre
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, Australia.
authors.forename: Arnan
authors.initials: A
authors.lastname: Mitchell
authors.affiliation: School of Electrical and Computer Engineering, RMIT University, Melbourne, Australia.
authors.forename: Khashayar
authors.initials: K
authors.lastname: Khoshmanesh
mesh_terms: D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology*; D002469:Cell Separation / Q000295:instrumentation*; D004586:Electrophoresis / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology; D056656:Lab-On-A-Chip Devices; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D050916:TRPV Cation Channels / Q000502:physiology*
chemical_list: D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; D002118:Calcium