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Method Article
Henning Gram Hansen
CHO Cell Line Engineering and Design, NNF-CfB, Technical University of Denmark
Corresponding Author
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This work is licensed under a CC BY-NC 3.0 License. Read Full License
Accurate assays for counting mammalian cells are important tools when performing experiments in cell biology. In general, commercially available cell-counting assays suffer from low throughput and high costs of consumables. Here, we describe a cost-efficient Hoechst and propidium iodide (PI)-staining protocol that accurately counts the number of viable mammalian cells in 96-well microplates based on image cytometry on a Celígo® Image Cytometer. Hands-on time for one microplate is less than 20 minutes and the entire procedure including incubation takes approximately 1 hour. Since all steps prior to image cytometry analysis involve liquid handling and since the Celígo® Image Cytometer is automation-friendly, this protocol has the potential to be integrated into automated liquid handling workflows.
Keywords
VCD, viability, mammalian cells, CHO, high-throughput, Celigo
Figure 1
I declare no conflict of interest.
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Associated Publications
Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells
by Henning Gram Hansen, Claes Nymand Nilsson, Anne Mathilde Lund, +7
Scientific Reports (21 March, 2016)
Method Article
Henning Gram Hansen
CHO Cell Line Engineering and Design, NNF-CfB, Technical University of Denmark
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 20 Jun, 2016
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Accurate assays for counting mammalian cells are important tools when performing experiments in cell biology. In general, commercially available cell-counting assays suffer from low throughput and high costs of consumables. Here, we describe a cost-efficient Hoechst and propidium iodide (PI)-staining protocol that accurately counts the number of viable mammalian cells in 96-well microplates based on image cytometry on a Celígo® Image Cytometer. Hands-on time for one microplate is less than 20 minutes and the entire procedure including incubation takes approximately 1 hour. Since all steps prior to image cytometry analysis involve liquid handling and since the Celígo® Image Cytometer is automation-friendly, this protocol has the potential to be integrated into automated liquid handling workflows.
Figure 1
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Associated Publications
Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells
by Henning Gram Hansen, Claes Nymand Nilsson, Anne Mathilde Lund, +7
Scientific Reports (21 March, 2016)