title: Image-based analysis of primary human neutrophil chemotaxis in an automated direct-viewing assay.
abstract: Multi-well assays based on the Boyden chamber have enabled highly parallel studies of chemotaxis-the directional migration of cells in response to molecular gradients-while direct-viewing approaches have allowed more detailed questions to be asked at low throughput. Boyden-based plates provide a count of cells that pass through a membrane, but no information about cell appearance. In contrast, direct-viewing devices enable the observation of cells during chemotaxis, which allows measurement of many parameters including area, shape, and location. Here we show automated chemotaxis and cell morphology assays in a 96-unit direct-viewing plate. Using only 12000 primary human neutrophils per datum, we measured dose-dependent stimulation and inhibition of chemotaxis and quantified the effects of inhibitors on cell area and elongation. With 60 parallel conditions we demonstrated 5-fold increase in throughput compared to previously reported direct-viewing approaches.
authors.affiliation: Bellbrook Labs, 5500 Nobel Dr., Ste. 250, Madison, WI 53711, USA. ivar.meyvantsson@bellbrooklabs.com
authors.forename: Ivar
authors.initials: I
authors.lastname: Meyvantsson
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Vu
authors.forename: Casey
authors.initials: C
authors.lastname: Lamers
authors.forename: Daniella
authors.initials: D
authors.lastname: Echeverria
authors.forename: Tracy
authors.initials: T
authors.lastname: Worzella
authors.forename: Victoria
authors.initials: V
authors.lastname: Echeverria
authors.forename: Allyson
authors.initials: A
authors.lastname: Skoien
authors.forename: Steven
authors.initials: S
authors.lastname: Hayes
mesh_terms: D002634:Chemotaxis, Leukocyte / Q000276:immunology*; D004867:Equipment Design; D006801:Humans; D007158:Immunologic Techniques; D016209:Interleukin-8 / Q000276:immunology; D046210:Microfluidic Analytical Techniques; D009504:Neutrophils / Q000166:cytology / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: C508606:CXCL8 protein, human; D016209:Interleukin-8; D011994:Recombinant Proteins