Multicolor plate reader fluorescence calibration

Jacob Beal*, Cheryl A. Telmer, Alejandro Vignoni, Yadira Boada, Geoff S. Baldwin, Liam Hallett, Taeyang Lee, Vinoo Selvarajah, Sonja Billerbeck, Bradley Brown, Guo Nan Cai, Liang Cai, Edward Eisenstein, Daisuke Kiga, David Ross, Nina Alperovich, Noah Sprent, Jaclyn Thompson, Eric M. Young, Drew EndyTraci Haddock-Angelli

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numberysac010
JournalSynthetic Biology
Volume7
Issue number1
DOIs
Publication statusPublished - 2022

Keywords

  • calibration
  • cell count
  • fluorescence
  • units

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomaterials
  • Biomedical Engineering
  • Agricultural and Biological Sciences (miscellaneous)

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