PicoQuant and the Charité – Universitätsmedizin Berlin are co-supervising an Early Stage Researcher (ESR) as part of the Marie Skłodowska-Curie actions in the Innovative Training Networks H2020-MSCA-ITN.
During his stay at PicoQuant, the ESR is preparing a PhD thesis on "Novel Applications for Spectrally Resolved Fluorescence Lifetime Imaging (sFLIM)".
The projects aim is to explore the feasibility and limits for quantitative separation of fluorescent markers in a strongly autofluorescing environment. The separation has been improved over the last years by using spectral confocal microscopy in combination with linear unmixing. However, the separation of multiple labels in biological samples still remains challenging, especially when strong tissue autofluorescence drowns out the response from specific labeled structures. By combining spectrally resolved detection with fluorescence lifetime measurements, simultaneous detection of spectral and lifetime parameters becomes possible, which is expected to significantly improve separation quality.
Early on in the research project, hard- and software parameters were established that are excellent for simultaneous spectrally resolved imaging and fluorescence lifetime measurement. In a cooperation with the Charité Berlin, the developed sFLIM detection set-up is used to investigate and analyze highly autofluorescent lung tissues that are labeled with multiple fluorophores. Applying a unique pattern matching technique1, a massive improved discrimination of these labels has been achieved.
1) Multi-target spectrally resolved fluorescence lifetime imaging microscopy, T. Niehörster et al., Nature Methods, 257-262, 13(3), 2016