Robust estimation of bacterial cell count from optical density

NTNU_Trondheim; NTU-Singapore; NU_Kazakhstan; NUDT_CHINA; NUS_Singapore-A; NUS_Singapore-Sci; NWU-China; NYMU-Taipei; NYU_Abu_Dhabi; OUC-China; Oxford; Paris_Bettencourt; Pasteur_Paris; Peking; Pittsburgh; Purdue; Queens_Canada; RDFZ-China; REC-CHENNAI; Rheda_Bielefeld; RHIT; Rice; Ruia-Mumbai; SBS_SH_112144; SCAU-China; SCU-China; SCUT_ChinaB; SCUT-ChinaA; SDU-CHINA; SFLS_Shenzhen; ShanghaiTech; SHSBNU_China; SHSID_China; SHSU_China; SIAT-SCIE; SJTU-BioX-Shanghai; SKLMT-China; SMMU-China; SMS_Shenzhen; Sorbonne_U_Paris; SSHS-Shenzhen; SSTi-SZGD; St_Andrews; Stanford; Stanford-Brown-RISD; Stockholm; Stony_Brook; Stuttgart; SUIS_Shanghai; SYSU-CHINA; SYSU-Software; SZU-China; Tacoma_RAINmakers; Tartu_TUIT; TAS_Taipei; Tec-Chihuahua; Tec-Monterrey; TecCEM; TecMonterrey_GDL; Thessaloniki; Tongji_China; Toronto; Toulouse-INSA-UPS; TU_Darmstadt; TU-Eindhoven; TUDelft; TUST_China; UAlberta; UC_Davis; UC_San_Diego; UCAS-China; UChicago; UChile_Biotec; UCL; UCLouvain; UConn; UCopenhagen; UESTC-China; UFlorida; UGA; UI_Indonesia; UiOslo_Norway; UIOWA; UIUC_Illinois; ULaval; ULaVerne_Collab; UMaryland; UNebraska-Lincoln; Unesp_Brazil; UNSW_Australia; UPF_CRG_Barcelona; UPF_CRG_Barcelona; US_AFRL_CarrollHS; USMA-West_Point; USP-Brazil; UST_Beijing; USTC; UT-Knoxville; Utrecht; Valencia_UPV; Vilnius-Lithuania; Vilnius-Lithuania-OG; Virginia; Warwick; Washington; WashU_StLouis; Waterloo; Westminster_UK; WHU-China; William_and_Mary; Worldshaper-Wuhan; WPI_Worcester; XJTLU-CHINA; XJTU-China; XMU-China; Yale; ZJU-China; ZJUT-China; iGEM Interlab Study Contributors; Aachen; Aalto-Helsinki; AHUT_China; Aix-Marseille; ASTWS-China; Athens; Austin_LASA; Austin_UTexas; Baltimore_BioCrew; BCU; BFSUICC-China; BGIC-Global; BGU_Israel; Bielefeld-CeBiTec; Bilkent-UNAMBG; BioIQS-Barcelona; BioMarvel; BIT; BIT-China; BJRS_China; BNDS_CHINA; BNU-China; BOKU-Vienna; BostonU; British_Columbia; Calgary; Cardiff_Wales; CCU_Taiwan; CDHSU-CHINA; Chalmers-Gothenburg; CIEI-BJ; CMUQ; CO_Mines; ColumbiaNYC; Cornell; CPU_CHINA; CSU_CHINA; CSU_Fort_Collins; Delgado-Ivy-Marin; DLUT_China; DLUT_China_B; DNHS_SanDiego; DTU-Denmark; Duesseldorf; Ecuador; ECUST; Edinburgh_OG; Edinburgh_UG; Emory; EPFL; ETH_Zurich; Evry_Paris-Saclay; Exeter; FAU_Erlangen; FJNU-China; Fudan; Fudan-CHINA; GDSYZX; Georgia_State; Gifu; GO_Paris-Saclay; GreatBay_China; Groningen; GZHS-United; HAFS; Hamburg; HBUT-China; HebrewU; HFLS_ZhejiangUnited; HK_HCY_LFC; HKJS_S; Hong_Kong_HKU; Hong_Kong_HKUST; Hong_Kong_JSS; Hong_Kong-CUHK; HUBU-Wuhan; HUST-China; HZAU-China; ICT-Mumbai; IISc-Bangalore; IISER-Bhopal-India; IISER-Kolkata; IISER-Mohali; IIT_Delhi; IIT_Kanpur; IIT-Madras; Jiangnan_China; Jilin_China; JMU_Wuerzburg; KAIT_JAPAN; KCL_UK; KUAS_Korea; Lambert_GA; Leiden; Lethbridge; Lethbridge_HS; Lubbock_TTU; Lund; Macquarie_Australia; Madrid-OLM; Manchester; Marburg; McGill; McMaster; METU_HS_Ankara; Michigan; MichiganState; Mingdao; Minnesota; Montpellier; Munich; Nanjing-China; NAU-CHINA; NAWI_Graz; NCHU_Taichung; NCTU_Formosa; NEU_China_A; NEU_China_B; Newcastle; NJU-China; NKU_CHINA; Northwestern; Nottingham; NPU-China; NTHU_Formosa; NTHU_Taiwan

doi:10.1038/s42003-020-01127-5

Robust estimation of bacterial cell count from optical density

NTNU_Trondheim, NTU-Singapore, NU_Kazakhstan, NUDT_CHINA, NUS_Singapore-A, NUS_Singapore-Sci, NWU-China, NYMU-Taipei, NYU_Abu_Dhabi, OUC-China, Oxford, Paris_Bettencourt, Pasteur_Paris, Peking, Pittsburgh, Purdue, Queens_Canada, RDFZ-China, REC-CHENNAI, Rheda_BielefeldRHIT, Rice, Ruia-Mumbai, SBS_SH_112144, SCAU-China, SCU-China, SCUT_ChinaB, SCUT-ChinaA, SDU-CHINA, SFLS_Shenzhen, ShanghaiTech, SHSBNU_China, SHSID_China, SHSU_China, SIAT-SCIE, SJTU-BioX-Shanghai, SKLMT-China, SMMU-China, SMS_Shenzhen, Sorbonne_U_Paris, SSHS-Shenzhen, SSTi-SZGD, St_Andrews, Stanford, Stanford-Brown-RISD, Stockholm, Stony_Brook, Stuttgart, SUIS_Shanghai, SYSU-CHINA, SYSU-Software, SZU-China, Tacoma_RAINmakers, Tartu_TUIT, TAS_Taipei, Tec-Chihuahua, Tec-Monterrey, TecCEM, TecMonterrey_GDL, Thessaloniki, Tongji_China, Toronto, Toulouse-INSA-UPS, TU_Darmstadt, TU-Eindhoven, TUDelft, TUST_China, UAlberta, UC_Davis, UC_San_Diego, UCAS-China, UChicago, UChile_Biotec, UCL, UCLouvain, UConn, UCopenhagen, UESTC-China, UFlorida, UGA, UI_Indonesia, UiOslo_Norway, UIOWA, UIUC_Illinois, ULaval, ULaVerne_Collab, UMaryland, UNebraska-Lincoln, Unesp_Brazil, UNSW_Australia, UPF_CRG_Barcelona, UPF_CRG_Barcelona, US_AFRL_CarrollHS, USMA-West_Point, USP-Brazil, UST_Beijing, USTC, UT-Knoxville, Utrecht, Valencia_UPV, Vilnius-Lithuania, Vilnius-Lithuania-OG, Virginia, Warwick, Washington, WashU_StLouis, Waterloo, Westminster_UK, WHU-China, William_and_Mary, Worldshaper-Wuhan, WPI_Worcester, XJTLU-CHINA, XJTU-China, XMU-China, Yale, ZJU-China, ZJUT-China, iGEM Interlab Study Contributors, Aachen, Aalto-Helsinki, AHUT_China, Aix-Marseille, ASTWS-China, Athens, Austin_LASA, Austin_UTexas, Baltimore_BioCrew, BCU, BFSUICC-China, BGIC-Global, BGU_Israel, Bielefeld-CeBiTec, Bilkent-UNAMBG, BioIQS-Barcelona, BioMarvel, BIT, BIT-China, BJRS_China, BNDS_CHINA, BNU-China, BOKU-Vienna, BostonU, British_Columbia, Calgary, Cardiff_Wales, CCU_Taiwan, CDHSU-CHINA, Chalmers-Gothenburg, CIEI-BJ, CMUQ, CO_Mines, ColumbiaNYC, Cornell, CPU_CHINA, CSU_CHINA, CSU_Fort_Collins, Delgado-Ivy-Marin, DLUT_China, DLUT_China_B, DNHS_SanDiego, DTU-Denmark, Duesseldorf, Ecuador, ECUST, Edinburgh_OG, Edinburgh_UG, Emory, EPFL, ETH_Zurich, Evry_Paris-Saclay, Exeter, FAU_Erlangen, FJNU-China, Fudan, Fudan-CHINA, GDSYZX, Georgia_State, Gifu, GO_Paris-Saclay, GreatBay_China, Groningen, GZHS-United, HAFS, Hamburg, HBUT-China, HebrewU, HFLS_ZhejiangUnited, HK_HCY_LFC, HKJS_S, Hong_Kong_HKU, Hong_Kong_HKUST, Hong_Kong_JSS, Hong_Kong-CUHK, HUBU-Wuhan, HUST-China, HZAU-China, ICT-Mumbai, IISc-Bangalore, IISER-Bhopal-India, IISER-Kolkata, IISER-Mohali, IIT_Delhi, IIT_Kanpur, IIT-Madras, Jiangnan_China, Jilin_China, JMU_Wuerzburg, KAIT_JAPAN, KCL_UK, KUAS_Korea, Lambert_GA, Leiden, Lethbridge, Lethbridge_HS, Lubbock_TTU, Lund, Macquarie_Australia, Madrid-OLM, Manchester, Marburg, McGill, McMaster, METU_HS_Ankara, Michigan, MichiganState, Mingdao, Minnesota, Montpellier, Munich, Nanjing-China, NAU-CHINA, NAWI_Graz, NCHU_Taichung, NCTU_Formosa, NEU_China_A, NEU_China_B, Newcastle, NJU-China, NKU_CHINA, Northwestern, Nottingham, NPU-China, NTHU_Formosa, NTHU_Taiwan

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

128 Scopus citations

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.

Original language	English
Article number	512
Journal	Communications Biology
Volume	3
Issue number	1
DOIs	https://doi.org/10.1038/s42003-020-01127-5
State	Published - 1 Dec 2020

ASJC Scopus subject areas

Medicine (miscellaneous)
General Biochemistry, Genetics and Molecular Biology
General Agricultural and Biological Sciences

Access to Document

10.1038/s42003-020-01127-5

Cite this

NTNU_Trondheim, NTU-Singapore, NU_Kazakhstan, NUDT_CHINA, NUS_Singapore-A, NUS_Singapore-Sci, NWU-China, NYMU-Taipei, NYU_Abu_Dhabi, OUC-China, Oxford, Paris_Bettencourt, Pasteur_Paris, Peking, Pittsburgh, Purdue, Queens_Canada, RDFZ-China, REC-CHENNAI, ... NTHU_Taiwan (2020). Robust estimation of bacterial cell count from optical density. Communications Biology, 3(1), Article 512. https://doi.org/10.1038/s42003-020-01127-5

@article{3cb4dcd34c7e4b779a682ff3c09f6056,

title = "Robust estimation of bacterial cell count from optical density",

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.",

author = "NTNU_Trondheim and NTU-Singapore and NU_Kazakhstan and NUDT_CHINA and NUS_Singapore-A and NUS_Singapore-Sci and NWU-China and NYMU-Taipei and NYU_Abu_Dhabi and OUC-China and Oxford and Paris_Bettencourt and Pasteur_Paris and Peking and Pittsburgh and Purdue and Queens_Canada and RDFZ-China and REC-CHENNAI and Rheda_Bielefeld and RHIT and Rice and Ruia-Mumbai and SBS_SH_112144 and SCAU-China and SCU-China and SCUT_ChinaB and SCUT-ChinaA and SDU-CHINA and SFLS_Shenzhen and ShanghaiTech and SHSBNU_China and SHSID_China and SHSU_China and SIAT-SCIE and SJTU-BioX-Shanghai and SKLMT-China and SMMU-China and SMS_Shenzhen and Sorbonne_U_Paris and SSHS-Shenzhen and SSTi-SZGD and St_Andrews and Stanford and Stanford-Brown-RISD and Stockholm and Stony_Brook and Stuttgart and SUIS_Shanghai and SYSU-CHINA and SYSU-Software and SZU-China and Tacoma_RAINmakers and Tartu_TUIT and TAS_Taipei and Tec-Chihuahua and Tec-Monterrey and TecCEM and TecMonterrey_GDL and Thessaloniki and Tongji_China and Toronto and Toulouse-INSA-UPS and TU_Darmstadt and TU-Eindhoven and TUDelft and TUST_China and UAlberta and UC_Davis and UC_San_Diego and UCAS-China and UChicago and UChile_Biotec and UCL and UCLouvain and UConn and UCopenhagen and UESTC-China and UFlorida and UGA and UI_Indonesia and UiOslo_Norway and UIOWA and UIUC_Illinois and ULaval and ULaVerne_Collab and UMaryland and UNebraska-Lincoln and Unesp_Brazil and UNSW_Australia and UPF_CRG_Barcelona and UPF_CRG_Barcelona and US_AFRL_CarrollHS and USMA-West_Point and USP-Brazil and UST_Beijing and USTC and UT-Knoxville and Utrecht and Valencia_UPV and Vilnius-Lithuania and Vilnius-Lithuania-OG and Virginia and Warwick and Washington and WashU_StLouis and Waterloo and Westminster_UK and WHU-China and William_and_Mary and Worldshaper-Wuhan and WPI_Worcester and XJTLU-CHINA and XJTU-China and XMU-China and Yale and ZJU-China and ZJUT-China and {iGEM Interlab Study Contributors} and Aachen and Aalto-Helsinki and AHUT_China and Aix-Marseille and ASTWS-China and Athens and Austin_LASA and Austin_UTexas and Baltimore_BioCrew and BCU and BFSUICC-China and BGIC-Global and BGU_Israel and Bielefeld-CeBiTec and Bilkent-UNAMBG and BioIQS-Barcelona and BioMarvel and BIT and BIT-China and BJRS_China and BNDS_CHINA and BNU-China and BOKU-Vienna and BostonU and British_Columbia and Calgary and Cardiff_Wales and CCU_Taiwan and CDHSU-CHINA and Chalmers-Gothenburg and CIEI-BJ and CMUQ and CO_Mines and ColumbiaNYC and Cornell and CPU_CHINA and CSU_CHINA and CSU_Fort_Collins and Delgado-Ivy-Marin and DLUT_China and DLUT_China_B and DNHS_SanDiego and DTU-Denmark and Duesseldorf and Ecuador and ECUST and Edinburgh_OG and Edinburgh_UG and Emory and EPFL and ETH_Zurich and Evry_Paris-Saclay and Exeter and FAU_Erlangen and FJNU-China and Fudan and Fudan-CHINA and GDSYZX and Georgia_State and Gifu and GO_Paris-Saclay and GreatBay_China and Groningen and GZHS-United and HAFS and Hamburg and HBUT-China and HebrewU and HFLS_ZhejiangUnited and HK_HCY_LFC and HKJS_S and Hong_Kong_HKU and Hong_Kong_HKUST and Hong_Kong_JSS and Hong_Kong-CUHK and HUBU-Wuhan and HUST-China and HZAU-China and ICT-Mumbai and IISc-Bangalore and IISER-Bhopal-India and IISER-Kolkata and IISER-Mohali and IIT_Delhi and IIT_Kanpur and IIT-Madras and Jiangnan_China and Jilin_China and JMU_Wuerzburg and KAIT_JAPAN and KCL_UK and KUAS_Korea and Lambert_GA and Leiden and Lethbridge and Lethbridge_HS and Lubbock_TTU and Lund and Macquarie_Australia and Madrid-OLM and Manchester and Marburg and McGill and McMaster and METU_HS_Ankara and Michigan and MichiganState and Mingdao and Minnesota and Montpellier and Munich and Nanjing-China and NAU-CHINA and NAWI_Graz and NCHU_Taichung and NCTU_Formosa and NEU_China_A and NEU_China_B and Newcastle and NJU-China and NKU_CHINA and Northwestern and Nottingham and NPU-China and NTHU_Formosa and NTHU_Taiwan and Jacob Beal and Farny, {Natalie G.} and Traci Haddock-Angelli and Vinoo Selvarajah and Baldwin, {Geoff S.} and Russell Buckley-Taylor and Markus Gershater and Daisuke Kiga and John Marken and Vishal Sanchania and Abigail Sison and Workman, {Christopher T.} and Meryem Pehlivan and Roige, {Biel Badia} and Tiu Aarnio and Samu Kivisto and Jessica Koski and Leevi Lehtonen and Denise Pezzutto and Pauliina Rautanen and Weixin Bian and Zhiyuan Hu and Zhihao Liu and Zi Liu and Liang Ma and Luyao Pan and Zichen Qin and Huichao Wang and Xiangxuan Wang and Hao Xu and Xia Xu and {El Moubayed}, Yorgo and Shan Dong and Choco Fang and Hanker He and Henry He and Fangliang Huang and Ruyi Shi and Cassie Tang and Christian Tang and Shirly Xu and Calvin Yan and Natalia Bartzoka and Eleni Kanata and Maria Kapsokefalou and Katopodi, {Xanthi Leda} and Eleni Kostadima and Kostopoulos, {Ioannis V.} and Dror Aizik and Mor Pasi",

note = "Publisher Copyright: {\textcopyright} 2020, The Author(s).",

year = "2020",

month = dec,

day = "1",

doi = "10.1038/s42003-020-01127-5",

language = "English",

volume = "3",

journal = "Communications Biology",

issn = "2399-3642",

publisher = "Springer Nature",

number = "1",

}

NTNU_Trondheim, NTU-Singapore, NU_Kazakhstan, NUDT_CHINA, NUS_Singapore-A, NUS_Singapore-Sci, NWU-China, NYMU-Taipei, NYU_Abu_Dhabi, OUC-China, Oxford, Paris_Bettencourt, Pasteur_Paris, Peking, Pittsburgh, Purdue, Queens_Canada, RDFZ-China, REC-CHENNAI, Rheda_Bielefeld, RHIT, Rice, Ruia-Mumbai, SBS_SH_112144, SCAU-China, SCU-China, SCUT_ChinaB, SCUT-ChinaA, SDU-CHINA, SFLS_Shenzhen, ShanghaiTech, SHSBNU_China, SHSID_China, SHSU_China, SIAT-SCIE, SJTU-BioX-Shanghai, SKLMT-China, SMMU-China, SMS_Shenzhen, Sorbonne_U_Paris, SSHS-Shenzhen, SSTi-SZGD, St_Andrews, Stanford, Stanford-Brown-RISD, Stockholm, Stony_Brook, Stuttgart, SUIS_Shanghai, SYSU-CHINA, SYSU-Software, SZU-China, Tacoma_RAINmakers, Tartu_TUIT, TAS_Taipei, Tec-Chihuahua, Tec-Monterrey, TecCEM, TecMonterrey_GDL, Thessaloniki, Tongji_China, Toronto, Toulouse-INSA-UPS, TU_Darmstadt, TU-Eindhoven, TUDelft, TUST_China, UAlberta, UC_Davis, UC_San_Diego, UCAS-China, UChicago, UChile_Biotec, UCL, UCLouvain, UConn, UCopenhagen, UESTC-China, UFlorida, UGA, UI_Indonesia, UiOslo_Norway, UIOWA, UIUC_Illinois, ULaval, ULaVerne_Collab, UMaryland, UNebraska-Lincoln, Unesp_Brazil, UNSW_Australia, UPF_CRG_Barcelona, UPF_CRG_Barcelona, US_AFRL_CarrollHS, USMA-West_Point, USP-Brazil, UST_Beijing, USTC, UT-Knoxville, Utrecht, Valencia_UPV, Vilnius-Lithuania, Vilnius-Lithuania-OG, Virginia, Warwick, Washington, WashU_StLouis, Waterloo, Westminster_UK, WHU-China, William_and_Mary, Worldshaper-Wuhan, WPI_Worcester, XJTLU-CHINA, XJTU-China, XMU-China, Yale, ZJU-China, ZJUT-China, iGEM Interlab Study Contributors, Aachen, Aalto-Helsinki, AHUT_China, Aix-Marseille, ASTWS-China, Athens, Austin_LASA, Austin_UTexas, Baltimore_BioCrew, BCU, BFSUICC-China, BGIC-Global, BGU_Israel, Bielefeld-CeBiTec, Bilkent-UNAMBG, BioIQS-Barcelona, BioMarvel, BIT, BIT-China, BJRS_China, BNDS_CHINA, BNU-China, BOKU-Vienna, BostonU, British_Columbia, Calgary, Cardiff_Wales, CCU_Taiwan, CDHSU-CHINA, Chalmers-Gothenburg, CIEI-BJ, CMUQ, CO_Mines, ColumbiaNYC, Cornell, CPU_CHINA, CSU_CHINA, CSU_Fort_Collins, Delgado-Ivy-Marin, DLUT_China, DLUT_China_B, DNHS_SanDiego, DTU-Denmark, Duesseldorf, Ecuador, ECUST, Edinburgh_OG, Edinburgh_UG, Emory, EPFL, ETH_Zurich, Evry_Paris-Saclay, Exeter, FAU_Erlangen, FJNU-China, Fudan, Fudan-CHINA, GDSYZX, Georgia_State, Gifu, GO_Paris-Saclay, GreatBay_China, Groningen, GZHS-United, HAFS, Hamburg, HBUT-China, HebrewU, HFLS_ZhejiangUnited, HK_HCY_LFC, HKJS_S, Hong_Kong_HKU, Hong_Kong_HKUST, Hong_Kong_JSS, Hong_Kong-CUHK, HUBU-Wuhan, HUST-China, HZAU-China, ICT-Mumbai, IISc-Bangalore, IISER-Bhopal-India, IISER-Kolkata, IISER-Mohali, IIT_Delhi, IIT_Kanpur, IIT-Madras, Jiangnan_China, Jilin_China, JMU_Wuerzburg, KAIT_JAPAN, KCL_UK, KUAS_Korea, Lambert_GA, Leiden, Lethbridge, Lethbridge_HS, Lubbock_TTU, Lund, Macquarie_Australia, Madrid-OLM, Manchester, Marburg, McGill, McMaster, METU_HS_Ankara, Michigan, MichiganState, Mingdao, Minnesota, Montpellier, Munich, Nanjing-China, NAU-CHINA, NAWI_Graz, NCHU_Taichung, NCTU_Formosa, NEU_China_A, NEU_China_B, Newcastle, NJU-China, NKU_CHINA, Northwestern, Nottingham, NPU-China, NTHU_Formosa & NTHU_Taiwan 2020, 'Robust estimation of bacterial cell count from optical density', Communications Biology, vol. 3, no. 1, 512. https://doi.org/10.1038/s42003-020-01127-5

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T1 - Robust estimation of bacterial cell count from optical density

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AU - OUC-China

AU - Oxford

AU - Paris_Bettencourt

AU - Pasteur_Paris

AU - Peking

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AU - Purdue

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AU - Toulouse-INSA-UPS

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AU - TUDelft

AU - TUST_China

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AU - UC_Davis

AU - UC_San_Diego

AU - UCAS-China

AU - UChicago

AU - UChile_Biotec

AU - UCL

AU - UCLouvain

AU - UConn

AU - UCopenhagen

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AU - UGA

AU - UI_Indonesia

AU - UiOslo_Norway

AU - UIOWA

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AU - UMaryland

AU - UNebraska-Lincoln

AU - Unesp_Brazil

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AU - UPF_CRG_Barcelona

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AU - USMA-West_Point

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AU - UST_Beijing

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AU - XJTLU-CHINA

AU - XJTU-China

AU - XMU-China

AU - Yale

AU - ZJU-China

AU - ZJUT-China

AU - iGEM Interlab Study Contributors

AU - Aachen

AU - Aalto-Helsinki

AU - AHUT_China

AU - Aix-Marseille

AU - ASTWS-China

AU - Athens

AU - Austin_LASA

AU - Austin_UTexas

AU - Baltimore_BioCrew

AU - BCU

AU - BFSUICC-China

AU - BGIC-Global

AU - BGU_Israel

AU - Bielefeld-CeBiTec

AU - Bilkent-UNAMBG

AU - BioIQS-Barcelona

AU - BioMarvel

AU - BIT

AU - BIT-China

AU - BJRS_China

AU - BNDS_CHINA

AU - BNU-China

AU - BOKU-Vienna

AU - BostonU

AU - British_Columbia

AU - Calgary

AU - Cardiff_Wales

AU - CCU_Taiwan

AU - CDHSU-CHINA

AU - Chalmers-Gothenburg

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AU - CMUQ

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AU - Cornell

AU - CPU_CHINA

AU - CSU_CHINA

AU - CSU_Fort_Collins

AU - Delgado-Ivy-Marin

AU - DLUT_China

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AU - Beal, Jacob

AU - Farny, Natalie G.

AU - Haddock-Angelli, Traci

AU - Selvarajah, Vinoo

AU - Baldwin, Geoff S.

AU - Buckley-Taylor, Russell

AU - Gershater, Markus

AU - Kiga, Daisuke

AU - Marken, John

AU - Sanchania, Vishal

AU - Sison, Abigail

AU - Workman, Christopher T.

AU - Pehlivan, Meryem

AU - Roige, Biel Badia

AU - Aarnio, Tiu

AU - Kivisto, Samu

AU - Koski, Jessica

AU - Lehtonen, Leevi

AU - Pezzutto, Denise

AU - Rautanen, Pauliina

AU - Bian, Weixin

AU - Hu, Zhiyuan

AU - Liu, Zhihao

AU - Liu, Zi

AU - Ma, Liang

AU - Pan, Luyao

AU - Qin, Zichen

AU - Wang, Huichao

AU - Wang, Xiangxuan

AU - Xu, Hao

AU - Xu, Xia

AU - El Moubayed, Yorgo

AU - Dong, Shan

AU - Fang, Choco

AU - He, Hanker

AU - He, Henry

AU - Huang, Fangliang

AU - Shi, Ruyi

AU - Tang, Cassie

AU - Tang, Christian

AU - Xu, Shirly

AU - Yan, Calvin

AU - Bartzoka, Natalia

AU - Kanata, Eleni

AU - Kapsokefalou, Maria

AU - Katopodi, Xanthi Leda

AU - Kostadima, Eleni

AU - Kostopoulos, Ioannis V.

AU - Aizik, Dror

AU - Pasi, Mor

PY - 2020/12/1

Y1 - 2020/12/1

N2 - 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.

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.

UR - http://www.scopus.com/inward/record.url?scp=85091192744&partnerID=8YFLogxK

U2 - 10.1038/s42003-020-01127-5

DO - 10.1038/s42003-020-01127-5

M3 - Article

C2 - 32943734

AN - SCOPUS:85091192744

SN - 2399-3642

VL - 3

JO - Communications Biology

JF - Communications Biology

IS - 1

M1 - 512

ER -

Robust estimation of bacterial cell count from optical density

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