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When Wavelengths Collide: Bias in Cell Abundance Measurements Due to Expressed Fluorescent Proteins
Ariel Hecht1,2,3, Drew Endy1,3, Marc Salit1,2,3
1Joint Initiative for Metrology in Biology, Stanford, California 94305, United States.
Abstract:
The abundance of bacteria in liquid culture is commonly inferred by measuring optical density at 600 nm. Red fluorescent proteins (RFPs) can strongly absorb light at 600 nm. Increasing RFP expression can falsely inflate apparent cell density and lead to underestimations of mean per-cell fluorescence by up to 10%. Measuring optical density at 700 nm would allow estimation of cell abundance unaffected by the presence of nearly all fluorescent proteins.
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