In vitro characterization of Fluorescence by Unbound Excitation from Luminescence: broadening the scope of energy transfer

Autor: Alexandra D. Holland, Joe Dragavon, Abdessalem Rekiki, Spencer L. Shorte, Jean-Yves Tinevez, Florian Rückerl, Régis Tournebize
Přispěvatelé: Imagerie Dynamique (Plate-Forme) (PFID), Institut Pasteur [Paris] (IP), Pathogénie Microbienne Moléculaire, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), The authors would like to extend their gratitude for financial support from: Institut Carnot Pasteur Maladies Infectieuses (to A.D.H., J.D., A.R., R.T., S.L.S.), Pasteur Foundation of New York (to J.D.), Region Ile de France program DimMalInf (S.L.S., R.T.), Programme Inter Carnot Fraunhofer PICF 2011 MEMI-OP (F.R.), ANR Program Grandes Investissement de l’avenir Infrastructures Nationales en Biologie-Santé: France LifebioImaging (FLI) France Life Imaging (R.T., S.L.S.), France Bioimaging (J.D., S.L.S), Programme LabEx: Project Biologie Intégrative des Maladies Infectieuses Emergentes (IBEID), and Institut Pasteur, Paris., the authors would like to thank Marie-Anne Nicola of the Plate-Forme d’Imagerie Dynamique for technical support and assistance, Cindy Fevre who generated the antibodies and Samantha Blazquez who purified them., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), Tinevez, Jean-Yves, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Institut Pasteur [Paris], Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)
Rok vydání: 2013
Předmět:
Fluorophore
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
[SDV]Life Sciences [q-bio]
MESH: Klebsiella pneumoniae
Analytical chemistry
MESH: Algorithms
MESH: Luciferases
Renilla / chemistry
010402 general chemistry
FUEL
01 natural sciences
Resonance (particle physics)
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
chemistry.chemical_compound
CRET
Quantum Dots
Radiative transfer
Escherichia coli
Molecular Biology
MESH: Fluorescent Dyes / chemistry
030304 developmental biology
Fluorescent Dyes
Luciferases
Renilla
0303 health sciences
Chemistry
Biochemistry
Genetics and Molecular Biology(all)
MESH: Escherichia coli
Fluorescence
MESH: Quantum Dots / chemistry
0104 chemical sciences
Characterization (materials science)
[SDV.BIO] Life Sciences [q-bio]/Biotechnology
[SDV] Life Sciences [q-bio]
Klebsiella pneumoniae
Förster resonance energy transfer
Spectrometry
Fluorescence
Energy Transfer
Chemical physics
FRET
BRET
MESH: Energy Transfer
Luminescence
Excitation
MESH: Spectrometry
Fluorescence
Algorithms
Zdroj: Methods
Methods, 2014, 66 (2), pp.353-361. ⟨10.1016/j.ymeth.2013.09.005⟩
Methods, Elsevier, 2014, 66 (2), pp.353-361. ⟨10.1016/j.ymeth.2013.09.005⟩
ISSN: 1095-9130
1046-2023
DOI: 10.1016/j.ymeth.2013.09.005⟩
Popis: Energy transfer mechanisms represent the basis for an array of valuable tools to infer interactions in vitro and in vivo , enhance detection or resolve interspecies distances such as with resonance. Based upon our own previously published studies and new results shown here we present a novel framework describing for the first time a model giving a view of the biophysical relationship between Fluorescence by Unbound Excitation from Luminescence (FUEL), a conventional radiative excitation–emission process, and bioluminescence resonance energy transfer. We show here that in homogeneous solutions and in fluorophore-targeted bacteria, FUEL is the dominant mechanism responsible for the production of red-shifted photons. The minor resonance contribution was ascertained by comparing the intensity of the experimental signal to its theoretical resonance counterpart. Distinctive features of the in vitro FUEL signal include a macroscopic depth dependency, a lack of enhancement upon targeting at a constant fluorophore concentration c f and a non-square dependency on c f . Significantly, FUEL is an important, so far overlooked, component of all resonance phenomena which should guide the design of appropriate controls when elucidating interactions. Last, our results highlight the potential for FUEL as a means to enhance in vivo and in vitro detection through complex media while alleviating the need for targeting.
Databáze: OpenAIRE
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