Skip to Main content Skip to Navigation
Journal articles

Green autofluorescence, a double edged monitoring tool for bacterial growth and activity in micro-plates

Abstract : The intrinsic green autofluorescence of an Escherichia coli culture has long been overlooked and empirically corrected in green fluorescent protein (GFP) reporter experiments. We show here, by using complementary methods of fluorescence analysis and HPLC, that this autofluorescence, principally arise from the secreted flavins in the external media. The cells secrete roughly 10 times more than what they keep inside. We show next that the secreted flavin fluorescence can be used as a complementary method in measuring the cell concentration particularly when the classical method, based on optical density measure, starts to fail. We also demonstrate that the same external flavins limit the dynamical range of GFP quantification and can lead to a false interpretation of lower global dynamic range of expression than what really happens. In the end we evaluate different autofluorescence correction methods to extract the real GFP signal.
Complete list of metadatas

Cited literature [29 references]  Display  Hide  Download

https://hal.archives-ouvertes.fr/hal-01628395
Contributor : Irina Mihalcescu <>
Submitted on : Thursday, January 31, 2019 - 11:52:10 AM
Last modification on : Wednesday, July 15, 2020 - 9:28:03 AM
Document(s) archivé(s) le : Wednesday, May 1, 2019 - 5:03:20 PM

File

Autofluorescence_with_Suppleme...
Files produced by the author(s)

Identifiers

Collections

Citation

Irina Mihalcescu, Mathilde Van-Melle Gateau, Bernard Chelli, Corinne Pinel, Jean-Luc Ravanat. Green autofluorescence, a double edged monitoring tool for bacterial growth and activity in micro-plates. Physical Biology, Institute of Physics: Hybrid Open Access, 2015, 12 (6), ⟨10.1088/1478-3975/12/6/066016⟩. ⟨hal-01628395⟩

Share

Metrics

Record views

264

Files downloads

688