Robust estimation of bacterial cell count from optical density
Communications Biology, vol. 3, pp. 512
Abstract
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 E. coli. 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.
Authors 78
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Jacob Beal corresponding
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Raytheon BBN Technologies, Cambridge, MA, USA. jakebeal@ieee.org
Raytheon BBN Technologies, Cambridge, MA, USA
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Natalie G. Farny corresponding
Worcester Polytechnic Institute
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Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, MA, USA. nfarny@wpi.edu
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, MA, USA
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Traci Haddock-Angelli corresponding
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iGEM Foundation, Cambridge, MA, USA. traci@igem.org
iGEM Foundation, Cambridge, MA, USA
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iGEM Foundation, Cambridge, MA, USA
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Geoff S. Baldwin corresponding
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Department of Life Sciences and IC-Centre for Synthetic Biology, Imperial College London, London, UK. g.baldwin@imperial.ac.uk
Department of Life Sciences and IC-Centre for Synthetic Biology, Imperial College London, London, UK
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Department of Life Sciences and IC-Centre for Synthetic Biology, Imperial College London, London, UK
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Markus C. Gershater corresponding
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Synthace, London, UK. m.gershater@synthace.com
Synthace, London, UK
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Faculty of Science and Engineering, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan
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California Institute of Technology
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Department of Bioengineering, California Institute of Technology, Pasadena, CA, USA
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Synthace, London, UK
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iGEM Foundation, Cambridge, MA, USA
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Christopher Thomas Workman corresponding
Technical University of Denmark
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DTU-Bioengineering, Technical University of Denmark, Kongens Lyngby, Denmark. cwor@dtu.dk
DTU-Bioengineering, Technical University of Denmark, Kongens Lyngby, Denmark
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Meryem Pehlivan Aachen
University of Helsinki · RWTH Aachen University · Aalto University
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RWTH Aachen University, Aachen, Germany
Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
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Biel Badia Roigé Aachen
University of Helsinki · RWTH Aachen University · Aalto University
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RWTH Aachen University, Aachen, Germany
Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
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Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
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Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
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Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
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Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
Anhui University of Technology, Maanshan, AnHui, China
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Aalto University, Espoo, Finland
University of Helsinki, Helsinki, Finland
Anhui University of Technology, Maanshan, AnHui, China
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Nanjing University · Anhui University of Technology
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Anhui University of Technology, Maanshan, AnHui, China
Nanjing University, Nanjing, China
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Anhui University of Technology, Maanshan, AnHui, China
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Anhui University of Technology, Maanshan, AnHui, China
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Anhui University of Technology, Maanshan, AnHui, China
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Anhui University of Technology, Maanshan, AnHui, China
Stony Brook University, Stony Brook, NY, USA
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Anhui University of Technology
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Anhui University of Technology, Maanshan, AnHui, China
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Nanjing Agricultural University, Nanjing, China
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National Technical University of Athens · Athens University of Economics and Business
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Athens University of Economics and Business, Athens, Attiki, Greece
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AST Worldshaper, Hangzhou, China
Athens University of Economics and Business, Athens, Attiki, Greece
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AST Worldshaper, Hangzhou, China
Athens University of Economics and Business, Athens, Attiki, Greece
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AST Worldshaper, Hangzhou, China
Nanjing Agricultural University, Nanjing, China
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National Technical University of Athens, Athens, Attiki, Greece
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University of British Columbia, Vancouver, BC, Canada
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Athens University of Economics and Business, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
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Athens University of Economics and Business, Athens, Attiki, Greece
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Liberal Arts and Science Academy High School, Austin, TX, USA
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
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National Technical University of Athens · Athens University of Economics and Business
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
Liberal Arts and Science Academy High School, Austin, TX, USA
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National Technical University of Athens · Athens University of Economics and Business
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
Liberal Arts and Science Academy High School, Austin, TX, USA
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National Technical University of Athens · Athens University of Economics and Business
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
Liberal Arts and Science Academy High School, Austin, TX, USA
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National Technical University of Athens · Athens University of Economics and Business
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
Liberal Arts and Science Academy High School, Austin, TX, USA
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National Technical University of Athens · Athens University of Economics and Business · The University of Texas at Austin
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Athens University of Economics and Business, Athens, Attiki, Greece
National Technical University of Athens, Athens, Attiki, Greece
The University of Texas at Austin, Austin, TX, USA
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Liberal Arts and Science Academy High School, Austin, TX, USA
Baltimore Underground Science Space, Baltimore, MD, USA
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Liberal Arts and Science Academy High School, Austin, TX, USA
Baltimore Underground Science Space, Baltimore, MD, USA
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Liberal Arts and Science Academy High School, Austin, TX, USA
Baltimore Underground Science Space, Baltimore, MD, USA
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Liberal Arts and Science Academy High School, Austin, TX, USA
Baltimore Underground Science Space, Baltimore, MD, USA
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University of British Columbia
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Liberal Arts and Science Academy High School, Austin, TX, USA
University of British Columbia, Vancouver, BC, Canada
Baltimore Underground Science Space, Baltimore, MD, USA
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Liberal Arts and Science Academy High School, Austin, TX, USA
Baltimore Underground Science Space, Baltimore, MD, USA
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Beijing City University · The University of Texas at Austin
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South China Agricultural University · Beijing Foreign Studies University · Ben-Gurion University of the Negev · Nanjing University
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Beijing Foreign Studies University, Beijing, China
Nanjing University, Nanjing, China
South China Agricultural University, Guangzhou, China
Ben-Gurion University of the Negev, Beer-Shiva, Israel
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Beijing Foreign Studies University, Beijing, China
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Shanghai Qibao Dwight High School, Shanghai, China
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Anhui University of Technology, Maanshan, AnHui, China
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Fudan University, Shanghai, China
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Institut Quimic de Sarria, Barcelona, Spain
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Ben-Gurion University of the Negev, Beer-Shiva, Israel
Institut Quimic de Sarria, Barcelona, Spain
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