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Cytotechnology

, Volume 68, Issue 5, pp 2015–2025 | Cite as

A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry

  • Leo Li-Ying Chan
  • Tim Smith
  • Kendra A. Kumph
  • Dmitry Kuksin
  • Sarah Kessel
  • Olivier Déry
  • Scott Cribbes
  • Ning Lai
  • Jean Qiu
Original Article

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.

Keywords

Image cytometry High-throughput Screening assay Viability Concentration Acridine orange Propidium iodide Celigo 

Notes

Compliance with ethical standards

Conflict of interest

The authors, LLC, TS, KK, DK, SK, OD, SC, NL, and JQ declare competing financial interests. The work performed in this manuscript is for reporting on product performance of Nexcelom Bioscience, LLC. The performed experiments were to demonstrate novel high-throughput screening method for cell concentration and viability using AO/PI on Celigo image cytometer.

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Leo Li-Ying Chan
    • 1
  • Tim Smith
    • 1
  • Kendra A. Kumph
    • 1
  • Dmitry Kuksin
    • 1
  • Sarah Kessel
    • 1
  • Olivier Déry
    • 1
  • Scott Cribbes
    • 1
  • Ning Lai
    • 1
  • Jean Qiu
    • 1
  1. 1.Department of Technology R&DNexcelom Bioscience LLCLawrenceUSA

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