期刊
SYNTHETIC BIOLOGY
卷 7, 期 1, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/synbio/ysac010
关键词
calibration; units; fluorescence; cell count
资金
- Air Force Research Laboratory (AFRL)
- DARPA [FA8750-17-C-0184]
- BBSRC [BB/M011178/1]
- EPSRC [EP/R034915/1, EP/S022856/1]
- YF Capital
- National Top Talent Undergraduate Training Program, China
- MINECO/AEI, EU [DPI2017-82896-C2-1-R]
- MCIN/AEI [PID2020117271RB-C21]
- Secretaria de Educacion Superior, Ciencia, Tecnologia e Innovacion-Ecuador
- JST, CREST, Japan [JPMJCR21N4]
- National Science Foundation [1939860]
- Office of the Director of National Intelligence (ODNI), Intelligence Advanced Research Projects Activity (IARPA) [N66001-18-C-4507]
- Office of Advanced Cyberinfrastructure (OAC)
- Direct For Computer & Info Scie & Enginr [1939860] Funding Source: National Science Foundation
This study extends the previous calibration protocols to include red fluorescence with sulforhodamine-101 fluorescent dye, and blue fluorescence with Cascade Blue. The interlaboratory study shows that these calibrants provide comparable precision to the previous ones and enable effective cross-laboratory comparison of red and blue fluorescence measurements.
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.
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