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PLATERO: A calibration protocol for plate reader green fluorescence measurements
Alba González-Cebrián1, Joan Borràs-Ferrís1, Yadira Boada2
1Multivariate Statistical Engineering Group, Department of Applied Statistics and O.R. and Quality, Universitat Politècnica de València, València, Spain.
Frontiers in Bioengineering and Biotechnology
|February 6, 2023
Summary
This study introduces PLATERO, a calibration protocol for synthetic biology fluorescence measurements. PLATERO standardizes data into concentration units, enabling reliable comparisons across experiments and devices.
Area of Science:
- Synthetic Biology
- Biophysics
- Biotechnology
Background:
- Plate reader fluorescence measurements are crucial in synthetic biology.
- Current measurements use arbitrary units and are affected by device gain, limiting comparability.
- Standardization is needed for accurate data analysis and reproducibility.
Purpose of the Study:
- To develop a calibration protocol (PLATERO) for standardizing fluorescence measurements.
- To express fluorescence intensity in concentration units of a reference fluorophore.
- To enable gain-independent and comparable fluorescence data.
Main Methods:
- Developed PLATERO, a calibration protocol using a Fluorescein model.
- Mathematical expression to normalize fluorescence data, removing device gain effects.
- Integrated Linearity, Bias, Reproducibility, and Repeatability analyses.
Main Results:
- PLATERO provides normalized, gain-independent fluorescence data.
- Enables comparison of measurements acquired at different gain levels.
- Quantifies uncertainty and variability in fluorescence measurements.
Conclusions:
- PLATERO offers a robust method for standardizing fluorescence data in synthetic biology.
- Enhances data comparability, reproducibility, and reliability.
- Associated functions are available in an open-access repository.
