Single cell clonal screening an important step in monoclonal antibody production cell line optimization, stable cell line construction, gene editing, and drug target screening.

With the xCELLigence RTCA eSight Dilution Cloning assays, you can now use whole well imaging (5x objective) to quickly identify colonies and verify monoclonality. With automated imaging and software, easily pinpoint clones derived from a single cell and track colony formation in real-time. Using either label or label-free methods, researchers can easily measure colony growth and monitor growth characteristics of clones (i.e. rapidly dividing). Instead of relying on labor intensive and manual-based methods that can be subject to error, the eSight allows for rapid identification of wells that contain clones of interest– all inside your incubator.

Using plate view of colonies over time (i.e. Day 0 to Day 10) eSight enables and provides simple and accurate screening of monoclonal cell lines with insightful data. The tracking and monitoring of monoclonal formation is made fully possible using the Dilution Cloning eSight software module, and brightfield and fluorescent focusing. Confluency and object count metrics allow you to quantify emerging colonies, while having the flexibility to stop, rewind, or move forward based on points of interest.

 


Through consecutive 10-fold dilutions, the cell suspension density is adjusted.
(Part 1) Tracking and monitoring of monoclonal formation using the brightfield focusing of Dilution Cloning module on RTCA eSight system. Representative images show the entire growth process from single cell to a clone (recognized cells are masked in orange) and the eSight system is able to recognize the single cell accurately.
(Part 2) Images of monoclonal formation collected using fluorescent focusing; the red object count is recognized by eSight software shown as a function of time. Live cell analysis reveals that only one cell was initially present in this particular well (from start), and clonal cluster growth was derived from one cell.

For more information regarding quantification and data output, please see further reading (literature) at: Agilent.com/chem/esight