title: Label-free imaging to study phenotypic behavioural traits of cells in complex co-cultures.
abstract: Time-lapse imaging is a fundamental tool for studying cellular behaviours, however studies of primary cells in complex co-culture environments often requires fluorescent labelling and significant light exposure that can perturb their natural function over time. Here, we describe ptychographic phase imaging that permits prolonged label-free time-lapse imaging of microglia in the presence of neurons and astrocytes, which better resembles in vivo microenvironments. We demonstrate the use of ptychography as an assay to study the phenotypic behaviour of microglial cells in primary neuronal co-cultures through the addition of cyclosporine A, a potent immune-modulator.
authors.affiliation: Phasefocus Ltd, Sheffield, UK.|Technology Facility, University of York, UK.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Suman
authors.affiliation: Department of Biology, University of York, UK.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Smith
authors.affiliation: Department of Biology, University of York, UK.
authors.forename: Kathryn E A
authors.initials: KE
authors.lastname: Hazel
authors.affiliation: Phasefocus Ltd, Sheffield, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Kasprowicz
authors.affiliation: Centre for Immunology and Infection, York, UK.|Department of Biology, University of York, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Coles
authors.affiliation: Technology Facility, University of York, UK.|Department of Biology, University of York, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: O'Toole
authors.affiliation: Department of Biology, University of York, UK.
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Chawla
mesh_terms: D000818:Animals; D018920:Coculture Techniques; D017628:Microglia; D009474:Neurons; D051381:Rats; D059008:Time-Lapse Imaging / Q000379:methods*