Dated: Jan 12, 2021
Publication Name: Molecular Devices
Citations : Rapid Monoclonality Verification Methods to Boost Cell Line Development
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.
Contributors: Wilson Lew, Anna Forsyth, John Philips, Alison Glaser, Natalia Lysaya