title: Sensitive and accurate quantification of human leukocyte migration using high-content Discovery-1 imaging system and ATPlite assay.
abstract: Migration is a fundamental aspect of leukocyte behavior and represents a significant therapeutic target clinically. However, most migration assays used in research are relatively low throughput and not easily compatible with rapid analysis or high-throughput screening (HTS) protocols required for drug screening assays. We therefore investigated the quantification of the migration of human leukocytes using the Molecular Devices high-content Discovery-1 platform or PerkinElmer ATPlite assay compared to manual counting. We have conducted extensive assay validation, investigating the detection limits, sensitivity, and precision of each method to count human leukocytes. Leukocyte migration assays were conducted using 96-well HTS-Transwell plates and the potent chemokine stromal cell-derived factor-1 (SDF-1). We reveal that the Discovery-1 and ATPlite methods developed here provide useful approaches to quantify leukocyte migration in an HTS manner with high levels of detection, sensitivity, and precision.
authors.affiliation: Centre for Brain Research and Department of Pharmacology and Clinical Pharmacology, School of Medical Sciences, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Natasha L
authors.initials: NL
authors.lastname: Grimsey
authors.forename: Kriebashne S
authors.initials: KS
authors.lastname: Moodley
authors.forename: Michelle
authors.initials: M
authors.lastname: Glass
authors.forename: E Scott
authors.initials: ES
authors.lastname: Graham
mesh_terms: D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D007118:Immunoassay; D007962:Leukocytes / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000032:analysis
chemical_list: D043562:Receptors, G-Protein-Coupled