Robust estimation of bacterial cell count from optical density

Jacob Beal, Natalie Farny, Traci Haddock-Angelli, Vinoo Selvarajah, Geoff Baldwin, Russell Buckley-Taylor, Markus Gershater, Daisuke Kiga, John Marken, Vishal Sanchania, Abigail Sison, Christopher T. Workman, iGEM Interlab Study Contributors, Aachen, Meryem Pehlivan, Biel Badia Roige, Aalto-Helsinki, Tiu Aarnio, Samu Kivisto, Jessica Koski, Leevi Lehtonen, Denise Pezzutto, Pauliina Rautanen, AHUT_China, Weixin Bian, Zhiyuan Hu, Zhihao Liu, Zi Liu, Liang Ma, Luyao Pan, Zichen Qin, Huichao Wang, Xiangxuan Wang, Hao Xu, Xia Xu, Aix-Marseille, Yorgo El Moubayed, ASTWS-China, Shan Dong, Choco Fang, Hanker He, Henry He, Fangliang Huang, Ruyi Shi, Cassie Tang, Christian Tang, Shirly Xu, Calvin Yan, Athens, Natalia Bartzoka, Eleni Kanata and Kyle Bowman 2020. Robust estimation of bacterial cell count from optical density. Communications Biology. 3 512. https://doi.org/10.1038/s42003-020-01127-5

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Title	Robust estimation of bacterial cell count from optical density
Type	Journal article
Authors	Jacob Beal, Natalie Farny, Traci Haddock-Angelli, Vinoo Selvarajah, Geoff Baldwin, Russell Buckley-Taylor, Markus Gershater, Daisuke Kiga, John Marken, Vishal Sanchania, Abigail Sison, Christopher T. Workman, iGEM Interlab Study Contributors, Aachen, Meryem Pehlivan, Biel Badia Roige, Aalto-Helsinki, Tiu Aarnio, Samu Kivisto, Jessica Koski, Leevi Lehtonen, Denise Pezzutto, Pauliina Rautanen, AHUT_China, Weixin Bian, Zhiyuan Hu, Zhihao Liu, Zi Liu, Liang Ma, Luyao Pan, Zichen Qin, Huichao Wang, Xiangxuan Wang, Hao Xu, Xia Xu, Aix-Marseille, Yorgo El Moubayed, ASTWS-China, Shan Dong, Choco Fang, Hanker He, Henry He, Fangliang Huang, Ruyi Shi, Cassie Tang, Christian Tang, Shirly Xu, Calvin Yan, Athens, Natalia Bartzoka, Eleni Kanata and Kyle Bowman
Abstract	<jats:title>Abstract</jats:title><jats:p>Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing <jats:italic>E. coli</jats:italic>. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.</jats:p>
Article number	512
Journal	Communications Biology
Journal citation	3
ISSN	2399-3642
Year	2020
Publisher	Springer Nature
Publisher's version	s42003-020-01127-5.pdf License CC BY 4.0 File Access Level Open (open metadata and files)
Digital Object Identifier (DOI)	https://doi.org/10.1038/s42003-020-01127-5
Web address (URL)	http://dx.doi.org/10.1038/s42003-020-01127-5
Published	17 Sep 2020

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Robust estimation of bacterial cell count from optical density

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