Journal article
Multicolor plate reader fluorescence calibration
Synthetic biology (Oxford University Press), Vol.7(1), pp.ysac010-ysac010
02/02/2022
DOI: 10.1093/synbio/ysac010
PMCID: PMC9357555
PMID: 35949424
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.
Details
- Title: Subtitle
- Multicolor plate reader fluorescence calibration
- Creators
- Jacob Beal - RTX (United States)Cheryl A. Telmer - Carnegie Mellon UniversityAlejandro Vignoni - Universitat Politècnica de ValènciaYadira Boada - Universitat Politècnica de ValènciaGeoff S. Baldwin - Imperial College LondonLiam Hallett - Imperial College LondonTaeyang Lee - Imperial College LondonVinoo Selvarajah - International Genetically Engineered Machine FoundationSonja Billerbeck - University of GroningenBradley Brown - Newcastle UniversityGuo-nan Cai - Fudan UniversityLiang Cai - Fudan UniversityEdward Eisenstein - Advanced Bioscience Laboratories (United States)Daisuke Kiga - Waseda UniversityDavid Ross - Material Measurement LaboratoryNina Alperovich - Material Measurement LaboratoryNoah Sprent - Imperial College LondonJaclyn Thompson - Worcester Polytechnic InstituteEric M. Young - Worcester Polytechnic InstituteDrew Endy - Stanford UniversityTraci Haddock-Angelli - International Genetically Engineered Machine Foundation
- Resource Type
- Journal article
- Publication Details
- Synthetic biology (Oxford University Press), Vol.7(1), pp.ysac010-ysac010
- DOI
- 10.1093/synbio/ysac010
- PMID
- 35949424
- PMCID
- PMC9357555
- NLM abbreviation
- Synth Biol (Oxf)
- ISSN
- 1939-7267
- eISSN
- 2397-7000
- Publisher
- Oxford Univ Press
- Number of pages
- 9
- Grant note
- PID2020117271RB-C21 / MCIN/AEI BB/M011178/1 / BBSRC; UK Research & Innovation (UKRI); Biotechnology and Biological Sciences Research Council (BBSRC) National Top Talent Undergraduate Training Program, China EP/R034915/1; EP/S022856/1 / EPSRC; UK Research & Innovation (UKRI); Engineering & Physical Sciences Research Council (EPSRC) DPI2017-82896-C2-1-R / MINECO/AEI, EU FA8750-17-C-0184 / DARPA; United States Department of Defense; Defense Advanced Research Projects Agency (DARPA) YF Capital 1939860 / National Science Foundation; National Science Foundation (NSF) Secretaria de Educacion Superior, Ciencia, Tecnologia e Innovacion-Ecuador N66001-18-C-4507 / Office of the Director of National Intelligence (ODNI), Intelligence Advanced Research Projects Activity (IARPA) Air Force Research Laboratory (AFRL); United States Department of Defense; US Air Force Research Laboratory JPMJCR21N4 / JST, CREST, Japan; Core Research for Evolutional Science and Technology (CREST)
- Language
- English
- Date published
- 02/02/2022
- Academic Unit
- Electrical and Computer Engineering
- Record Identifier
- 9984627235302771
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