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© Research
Publication : Communications biology

Robust estimation of bacterial cell count from optical density.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Communications biology - 17 Sep 2020

Beal J, Farny NG, Haddock-Angelli T, Selvarajah V, Baldwin GS, Buckley-Taylor R, Gershater M, Kiga D, Marken J, Sanchania V, Sison A, Workman CT; Deshmukh Gopaul as member of the iGEM Interlab Study Contributors.

Link to Pubmed [PMID] – 32943734

Link to DOI [DOI] – 10.1038/s42003-020-01127-5

Commun Biol 2020 Sep; 3(1): 512

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.

https://pubmed.ncbi.nlm.nih.gov/32943734