RT Journal Article
T1 Robust estimation of bacterial cell count from optical density
A1 Beal, Jacob
A1 Alonso Conde, Rafael Alejandro
A1 Hernández Cifuentes, Rodrigo
A1 Sánchez Clemente, Borja
A1 Saiz Gonzalo, Gonzalo
A1 Martín Hernández, Iván
A1 Armero Hernández, Laura
A1 Quero Lombardero, Francisco Javier
A1 Marquina Díaz, Domingo
A1 Fernández Rodríguez, Guillermo
A1 Albert Smet, Ignacio
A1 Zhou, Jiajie
AB 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.
PB Spring Nature
SN 2399-3642
YR 2020
FD 2020
LK https://hdl.handle.net/20.500.14352/107449
UL https://hdl.handle.net/20.500.14352/107449
LA eng
NO Beal, Jacob, et al. «Robust estimation of bacterial cell count from optical density». Communications Biology, vol. 3, n.º 1, 2020, https://doi.org/10.1038/S42003-020-01127-5.
NO NSF's Expeditions in Computing Awards
NO Engineering and Physical Sciences Research Council
NO Unión Europea
DS Docta Complutense
RD 27 abr 2025