Institut Pasteur

Kelly Rogers

Philippe Brulet

Spencer Shorte

Joseph Dragavon

Samantha Blazquez

Sandrine Picaud

Spencer L Shorte

The present invention relates a method to determine the presence of a photon producing biological marker in a cell, tissue or organism of interest. The method is based on Fluorescence by Unbound Excitation from Luminescence (FUEL) and comprises the steps of a) providing conditions suitable for the biological marker to produce at least one first photon by luminescence; b) providing a FUEL probe pair-upper (FPP-U) disposed in proximity to the cell, tissue or organism, wherein the at least one first photon of step a) excites the FPP-U, which emits at least one second photon. The FPP-U may be selected from the group of quantum dots, carbon nanotubes, fluorescent proteins, diamond nanocrystals and metalloporphyrins. This method is characterized in that said biological marker and said FPP-U are not bound and in that each of the at least one second photon(s) are of a longer wavelength than each of the at least one first photon(s).

Method to increase the number of detectable photons during the imaging of a biological marker

The invention relates to a method for optical detection of the dynamics of Cain a biological system, said method comprising monitoring the photons emitted by a recombinant Ca-sensitive polypeptide, which comprises or consists of a chemiluminescent protein linked to a fluorescent protein, present in said biological system. In a particular embodiment, said recombinant polypeptide comprises or consists of the Aequorin and GFP linked by a linker allowing chemiluminescence resonance energy transfer (CRET), and optionally comprises a peptidic fragment capable of targeting said recombinant polypeptide into a specific cellular domain or compartment. The present invention also concerns a transgenic non-human animal expressing said recombinant polypeptide sensitive to calcium concentration, in conditions enabling the in vivo monitoring of Cadynamics. In a particular embodiment, the expression and/or localization of said recombinant polypeptide is restricted to a specific tissue, a single-cell type and/or in a particular cellular compartment or domain.

Non-invasive real-time in vivo bioluminescence imaging of local Ca&#xc2;2+ dynamics in living organisms

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Victoria, Australia

Kelly Rogers