title: A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry.
abstract: To ensure cell-based assays are performed properly, both cell concentration and viability have to be determined so that the data can be normalized to generate meaningful and comparable results. Cell-based assays performed in immuno-oncology, toxicology, or bioprocessing research often require measuring of multiple samples and conditions, thus the current automated cell counter that uses single disposable counting slides is not practical for high-throughput screening assays. In the recent years, a plate-based image cytometry system has been developed for high-throughput biomolecular screening assays. In this work, we demonstrate a high-throughput AO/PI-based cell concentration and viability method using the Celigo image cytometer. First, we validate the method by comparing directly to Cellometer automated cell counter. Next, cell concentration dynamic range, viability dynamic range, and consistency are determined. The high-throughput AO/PI method described here allows for 96-well to 384-well plate samples to be analyzed in less than 7 min, which greatly reduces the time required for the single sample-based automated cell counter. In addition, this method can improve the efficiency for high-throughput screening assays, where multiple cell counts and viability measurements are needed prior to performing assays such as flow cytometry, ELISA, or simply plating cells for cell culture.
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA. lchan@nexcelom.com.
authors.forename: Leo Li-Ying
authors.initials: LL
authors.lastname: Chan
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Smith
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Kendra A
authors.initials: KA
authors.lastname: Kumph
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Kuksin
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Kessel
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Olivier
authors.initials: O
authors.lastname: Déry
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Cribbes
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Lai
authors.affiliation: Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack St. Building 9, Lawrence, MA, 01843, USA.
authors.forename: Jean
authors.initials: J
authors.lastname: Qiu
keywords: Acridine orange; Celigo; Concentration; High-throughput; Image cytometry; Propidium iodide; Screening assay; Viability