13 research outputs found
Development of chemiluminescent probes for the detection of enzymatic actvity
Depuis plusieurs annĂ©es, il est devenu plus aisĂ© de dĂ©tecter et dâĂ©tudier les mĂ©canismes biologiques in cellulo grĂące aux techniques dâimagerie optique que sont la fluorescence, la bioluminescence et la chimiluminescence. Bien que la fluorescence soit la technique la plus employĂ©e de nos jours, la bioluminescence et la chimiluminescence, qui sont la consĂ©quence dâune cascade de rĂ©actions (bio)chimiques, sont trĂšs Ă©tudiĂ©es depuis quelques dĂ©cennies. En effet, elles permettent de contourner la source des problĂšmes rencontrĂ©s dans lâutilisation dâun fluorophore : la lumiĂšre excitatrice. La chimiluminescence est par consĂ©quent une mĂ©thode de choix pour sâaffranchir de lâauto-fluorescence tissulaire. Le 1,2-dioxĂ©tane est un des motifs chimiluminescent qui, par dĂ©composition, peut gĂ©nĂ©rer un Ă©tat excitĂ© sur un fluorophore auquel il est connectĂ©, autorisant ainsi sa luminescence intrinsĂšque. Ainsi, le but de ce projet de thĂšse a Ă©tĂ© de dĂ©velopper et dâĂ©tudier des nouvelles sondes chĂ©miluminescentes Ă motif 1,2-dioxĂ©tane Ă coeurs naphtolique et phĂ©nolique, compatibles avec le vivant pour une dĂ©tection ciblĂ©e de complexes enzymatiques. La stratĂ©gie envisagĂ©e passait par la prĂ©paration d'une plateforme chimiluminescente comportant un motif 1,2-dioxĂ©tane thermiquement stable, sur laquelle il est possible de faire varier le dĂ©clencheur (i.e. possibilitĂ© d'adapter cette plateforme Ă l'analyte ou Ă©vĂ©nement que l'on souhaite dĂ©tecter) et d'accrocher un fluorophore NIR. Deux mĂ©thodes ont Ă©tĂ© tentĂ©es : dâune part une greffe dâune version hydrosoluble dâun fluorophore connu (i.e. le Nile red) pour rĂ©aliser un transfert dâĂ©nergie Ă travers les liaisons (i.e. TBET), et dâautre part un couplage Ă un complexe de lanthanide permettant un transfert dâĂ©nergie Ă travers lâespace (i.e. CRET).Since several years, it is easier to exploit biological phenomena in cellulo through technologies dealing with bioimaging. This method gathers fluorescence, bioluminescence and chemiluminescence. Even though fluorescence is the most employed technic, bioluminescence and chemiluminescence, being the consequence of (bio)chemical reaction, have been widely studied for decades. In fact, they can avoid the main problems encountered in fluorophore use: exciting light. Chemiluminescence is thus the appropriate approach to avoid biological autofluorescence. 1,2-dioxetane is one of the moieties that, upon decomposition, can generate an excited state on a connected fluorophore, giving rise to its intrinsic luminescence. The aim of our project was to develop and study new 1,2-dioxetan chemiluminescent probes based on phenol or naphthol moieties, for targeted enzymatic complexes detection with in cellulo and in vivo bioimaging. The strategy relied on the synthesis of chemiluminescent scaffolds comprising thermally stable 1,2-dioxetan, on which the trigger and the connected NIR fluorophore can be easily diversified. Two methods have been attempted: 1) coupling to a water-soluble version of a known fluorophore (i.e. Nile red) allowing an energy transfer through bonds (i.e. TBET) 2) connection with a lanthanide complex giving rise to an energy transfer through space (i.e. CRET)
Développement de sondes chimiluminescentes pour la détection d'activités enzymatiques
Since several years, it is easier to exploit biological phenomena in cellulo through technologies dealing with bioimaging. This method gathers fluorescence, bioluminescence and chemiluminescence. Even though fluorescence is the most employed technic, bioluminescence and chemiluminescence, being the consequence of (bio)chemical reaction, have been widely studied for decades. In fact, they can avoid the main problems encountered in fluorophore use: exciting light. Chemiluminescence is thus the appropriate approach to avoid biological autofluorescence. 1,2-dioxetane is one of the moieties that, upon decomposition, can generate an excited state on a connected fluorophore, giving rise to its intrinsic luminescence. The aim of our project was to develop and study new 1,2-dioxetan chemiluminescent probes based on phenol or naphthol moieties, for targeted enzymatic complexes detection with in cellulo and in vivo bioimaging. The strategy relied on the synthesis of chemiluminescent scaffolds comprising thermally stable 1,2-dioxetan, on which the trigger and the connected NIR fluorophore can be easily diversified. Two methods have been attempted: 1) coupling to a water-soluble version of a known fluorophore (i.e. Nile red) allowing an energy transfer through bonds (i.e. TBET) 2) connection with a lanthanide complex giving rise to an energy transfer through space (i.e. CRET).Depuis plusieurs annĂ©es, il est devenu plus aisĂ© de dĂ©tecter et dâĂ©tudier les mĂ©canismes biologiques in cellulo grĂące aux techniques dâimagerie optique que sont la fluorescence, la bioluminescence et la chimiluminescence. Bien que la fluorescence soit la technique la plus employĂ©e de nos jours, la bioluminescence et la chimiluminescence, qui sont la consĂ©quence dâune cascade de rĂ©actions (bio)chimiques, sont trĂšs Ă©tudiĂ©es depuis quelques dĂ©cennies. En effet, elles permettent de contourner la source des problĂšmes rencontrĂ©s dans lâutilisation dâun fluorophore : la lumiĂšre excitatrice. La chimiluminescence est par consĂ©quent une mĂ©thode de choix pour sâaffranchir de lâauto-fluorescence tissulaire. Le 1,2-dioxĂ©tane est un des motifs chimiluminescent qui, par dĂ©composition, peut gĂ©nĂ©rer un Ă©tat excitĂ© sur un fluorophore auquel il est connectĂ©, autorisant ainsi sa luminescence intrinsĂšque. Ainsi, le but de ce projet de thĂšse a Ă©tĂ© de dĂ©velopper et dâĂ©tudier des nouvelles sondes chĂ©miluminescentes Ă motif 1,2-dioxĂ©tane Ă coeurs naphtolique et phĂ©nolique, compatibles avec le vivant pour une dĂ©tection ciblĂ©e de complexes enzymatiques. La stratĂ©gie envisagĂ©e passait par la prĂ©paration d'une plateforme chimiluminescente comportant un motif 1,2-dioxĂ©tane thermiquement stable, sur laquelle il est possible de faire varier le dĂ©clencheur (i.e. possibilitĂ© d'adapter cette plateforme Ă l'analyte ou Ă©vĂ©nement que l'on souhaite dĂ©tecter) et d'accrocher un fluorophore NIR. Deux mĂ©thodes ont Ă©tĂ© tentĂ©es : dâune part une greffe dâune version hydrosoluble dâun fluorophore connu (i.e. le Nile red) pour rĂ©aliser un transfert dâĂ©nergie Ă travers les liaisons (i.e. TBET), et dâautre part un couplage Ă un complexe de lanthanide permettant un transfert dâĂ©nergie Ă travers lâespace (i.e. CRET)
Development of chemiluminescent probes for enzymatic activity detection.
International audienc
Development of chemiluminescent probes for enzymatic activity detection.
International audienc
Development of chemiluminescent probes for enzymatic activity detection.
International audienc
Design and Synthesis of Chemiluminescent Cassettes Based on Energy Transfer
International audienceChemiluminescent probes are being considered as a convenient option for optical imaging. Several strategies were reported to increase the probe chemiluminescence efficiency. In this study, a series of chemiluminescent cassettes based on adamantyl stabilized 1,2-dioxetanes ("Schaap's dioxetane") linked to a fluorophore (BODIPY or dicyanoisophorone fluorophore) by a conjugated linker have been synthetized. Their chemiluminescent decomposition and the photoluminescence properties of their respective emissive species were investigated
Development of chemiluminescent probes for enzymatic activity detection.
International audienc
Development of chemiluminescent probes for enzymatic activity detection.
International audienc