Multicolor plate reader fluorescence calibration-Reference-Cited by-同舟云学术

Multicolor plate reader fluorescence calibration

Published:2022-02-01 Issue:1 Volume:7 Page:
ISSN:2397-7000
Container-title:Synthetic Biology
language:en
Short-container-title:

Author:

Beal Jacob¹^ORCID,Telmer Cheryl A²^ORCID,Vignoni Alejandro³^ORCID,Boada Yadira³,Baldwin Geoff S⁴^ORCID,Hallett Liam⁴,Lee Taeyang⁴,Selvarajah Vinoo⁵,Billerbeck Sonja⁶^ORCID,Brown Bradley⁷,Cai Guo-nan⁸,Cai Liang⁸,Eisenstein Edward⁹,Kiga Daisuke¹⁰,Ross David¹¹^ORCID,Alperovich Nina¹¹,Sprent Noah¹²^ORCID,Thompson Jaclyn¹³,Young Eric M¹³,Endy Drew¹⁴,Haddock-Angelli Traci⁵

Affiliation:

1. Intelligent Software and Systems, Raytheon BBN Technologies , 10 Moulton Street, Cambridge 02138, MA, USA

2. Department of Biological Sciences, Carnegie Mellon University , 4400 Fifth Avenue, Pittsburgh 15213, PA, USA

3. Synthetic Biology and Biosystems Control Group, Instituto de Automatica e Informatica Industrial, Universitat Politecnica de Valencia , Camino de Vera s/n, Valencia 46022, Spain

4. Department of Life Sciences, Imperial College London , South Kensington Campus, Exhibition Road, London SW7 2AZ, UK

5. iGEM Foundation , 45 Prospect Street, Cambridge 02139, MA, USA

6. Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen , Nijenborgh 7, Groningen 9747 AG, The Netherlands

7. School of Engineering, Newcastle University , Devonshire Building, Devonshire Terrace, NE1 7RU Newcastle Upon Tyne, UK

8. School of Life Sciences, Fudan University , 220 Handan Road, Shanghai 200433, China

9. Fischell Department of Bioengineering, University of Maryland Institute of Bioscience and Biotechnology Research, , 9600 Gudelsky Drive, Rockville 20850, MD, USA

10. School of Advanced Science and Engineering, Waseda University , 2-2 Wakamatsu Cho, Totsukamachi, Shinjuku City 169-8050, Tokyo, Japan

11. Material Measurement Laboratory, National Institute of Standards and Technology , 100 Bureau Dr., Gaithersburg 20899, MD, USA

12. Department of Chemical Engineering, Imperial College London , South Kensington Campus, Exhibition Road, London SW7 2AZ, UK

13. Chemical Engineering, Worcester Polytechnic Institute , 100 Institute Road, Worcester 01609-2280, MA, USA

14. Bioengineering, Stanford University , 443 Via Ortega, Stanford 94305, CA, USA

Abstract

Abstract Plate readers are commonly used to measure cell growth and fluorescence, yet the utility and reproducibility of plate reader data is limited by the fact that it is typically reported in arbitrary or relative units. We have previously established a robust serial dilution protocol for calibration of plate reader measurements of absorbance to estimated bacterial cell count and for green fluorescence from proteins expressed in bacterial cells to molecules of equivalent fluorescein. We now extend these protocols to calibration of red fluorescence to the sulforhodamine-101 fluorescent dye and blue fluorescence to Cascade Blue. Evaluating calibration efficacy via an interlaboratory study, we find that these calibrants do indeed provide comparable precision to the prior calibrants and that they enable effective cross-laboratory comparison of measurements of red and blue fluorescence from proteins expressed in bacterial cells.

Funder

J.B.

N.S.

G.B., L.H., and T.L.

G.C. and L.C.

A.V. and Y.B.

Y.B.

JST, CREST

E.Y.

National Science Foundation

Publisher

Oxford University Press (OUP)