Citations : Clone Select Imager

A high-throughput, automated platform of manufacturing cell line development for producing protein therapeutics is described. Implementation of BD FACS Aria Cell Sorter, CloneSelect Imager and TECAN Freedom EVO liquid handling system has demonstrated significantly increased processing capacity in cell line development with improved cell line quality and high reproducibility.

Limiting dilution or fluorescence activated cell sorting is typically performed to seed single cells into a well. Microscopy is then used to determine the number of cells seeded in each well and monitor cell growth. While monoclonality verification via white light imaging is possible, debris, dust, and air bubbles make it difficult and time consuming to identify single cells which may cause high value clones to be discarded. Here we present a fluorescent method for identifying monoclonal CHO-S cells using CellTracker Green CMDFA. We incubated CHO-S cells with Cell Tracker Green CMDFA and performed limited dilution to seed single CHO-S cells into 96-well plates. The CloneSelect Imager was used to image CHO-S cells in white light and fluorescence channels. By using fluorescence to identify cells, monoclonality verification is easier and more conclusive than using white light imaging or microscopy alone.

High-throughput screening methods for toxicology evaluation in the early phases of drug discovery are highly desired. In the early 1980´s a novel assay for cell survival determination was reported [1], which has been frequently used to study the biological activity of a variety of potential cytostatic drugs. The assay was presented as a rapid, precise and simple method to detect living cells in mammalian cell cultures, using the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and a microtiter plate reader.