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Fluorescence-detected Transient Absorption Microscopy

Project description

Extending the power of transient-absorption spectroscopy

Transient-absorption spectroscopy is a powerful tool to monitor the dynamics of photo-excited states in a sample. Despite their potential, such ultrafast nonlinear techniques typically require large-volume samples and coherent detection. Funded by the Marie Skłodowska-Curie Actions programme, the FluoTRAM project will leverage a newly developed technique for detecting the fluorescence of a sample and apply it to transient-absorption spectroscopy. The technique should help reveal additional information on the excitation dynamics in the samples. This information could prove useful in many areas, including the correlation of the excitation and emission spectra for dye probes, intramolecular charge transfer in fluorescent proteins and charge transfer and recombination in organic materials.

Objective

Fluorescence microscopy is an indispensable tool in many areas of research. In life sciences it has been perfected for biological sample imaging either by its autofluorescence or using fluorescent markers such as dyes or fluorescent proteins. It is thus possible to localize molecules in cells, obtaining wealth of information on their dynamics and environment. Despite its power, the fluorescence detection is, by its nature, limited to the information on the final, emissive state of the molecules after photoexcitation. Meanwhile, transient absorption spectroscopy enables to track the initial state of the molecules after absorption and the following excitation dynamics. However, such ultrafast nonlinear techniques typically require volume samples and coherent detection. We have recently developed a new way to measure transient absorption by detecting the sample fluorescence. In project FluoTRAM we will implement our technique in the fluorescence microscope, where it truly reveals its potential. Using the established imaging techniques and markers, FluoTRAM brings the additional information on the excitation event and the dynamics towards the emissive state. We will implement FluoTRAM in two parallel stages, the time resolution and the spectrally varying excitation. The time resolution will be achieved using chopped laser pulses, varying their delay by a delay stage and recording a difference fluorescence in a pump-probe fashion. The excitation spectrum scanning will be realized interferometrically, creating a phase-stable replica of the excitation pulse and scanning the delay between the two. The comprehensive additional information on the excitation dynamics from absorption to emission will be of great use in life sciences and beyond. Examples include correlation of the excitation and emission spectra (increased Stokes shift vs red shift) for dye probes, intramolecular charge transfer in fluorescent proteins, or charge transfer and recombination in organic materials.

Coordinator

UNIVERZITA KARLOVA
Net EU contribution
€ 144 980,64
Address
OVOCNY TRH 560/5
116 36 Praha 1
Czechia

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Region
Česko Praha Hlavní město Praha
Activity type
Higher or Secondary Education Establishments
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Total cost
€ 144 980,64