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A Method for Quick, Low-Cost Automated Confluency Measurements

Published online by Cambridge University Press:  28 October 2011

Gil Topman ,
Orna Sharabani-Yosef  and
Amit Gefen
Gil Topman
Affiliation:
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv 69978, Israel
Orna Sharabani-Yosef
Affiliation:
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv 69978, Israel
Amit Gefen*
Affiliation:
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv 69978, Israel
*
Corresponding author. E-mail: gefen@eng.tau.ac.il
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Abstract

A culture's confluency is a fundamental measure in the field of biology, and routine quantification of confluence in cell culture protocols, biological assays and tissue engineering work is important. However, current techniques for obtaining confluency are either subjective, destructive, not simple enough, or time-consuming. We developed an image processing method for automated confluency measurement from a single microscope image without any chemical staining. To demonstrate utility we monitored the confluency of three cell types: NIH3T3 fibroblasts, C2C12 myoblasts, and 3T3L1 pre-adipocytes for 5 days, twice a day. The captured micrographs had different and uneven illumination, the cell types varied in cell-to-background contrast, and the confluency ranged between 10% and 100%. Despite these variable conditions, our method was shown to be practical, economic, and easy to implement, providing quantitative confluency measurements over time in each culture case. The method is hence suitable for routine automatic determination of confluency to standardize handling of cells, achieve reproducibility across trials, and improve accuracy in experimental outcome measures.

Keywords

image analysissegmentationtexturecell culturemicroscopytissue engineering
Type
Software and Techniques Development
Information
Microscopy and Microanalysis , Volume 17 , Issue 6 , December 2011 , pp. 915 - 922
Copyright
Copyright © Microscopy Society of America 2011

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References

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