Pushing Boundaries in FLIM to Enhance Efficiency, Quality and Reproducibility

14:30 – 15:30 BST, 6 June 2024 ‐ 1 hour

PicoQuant

Isabel Gross, Fabio Barachati, Marcelle Koenig, Maria Loidolt-Krueger, Ellen Schmeyer, Matthias Patting, Marcus Sackrow, Uwe Ortmann, Evangelos Sisamakis, Felix Koberling, Rainer Erdmann

Workshop Room 5

Quantitative time-resolved fluorescence techniques like Fluorescence Lifetime Imaging (FLIM) have become more attractive recently to study mechanisms driven by phase separation or to sense the cellular environment, for example. 

PicoQuant`s innovative confocal microscope Luminosa combines state-of-the-art hardware with cutting edge software to deliver high quality data while simplifying daily operation. The software includes several features which improve the ease of use and reproducibility of experiments, including context-based workflows, sample-free auto-alignment and excitation laser power calibration. Still, if required for new method development every optomechanical component can be fully accessible.

We will show how FLIM is streamlined with Luminosa. Luminosa’s rapidFLIM hardware can record several frames per second with high photon count rates, which the software handles with a novel dynamic binning format. In combination with GPU-accelerated algorithms, this enables high-speed automated analysis of FLIM images. The InstaFLIM analysis workflow suggests the best fitting model based on statistical arguments, requiring minimal user interaction. The optional NovaFLIM software package enables more extensive and advanced image analysis.

Many recent initiatives have focused their efforts on improving the aspects of Quality Assessment (QA), Quality Control (QC), and reproducibility in time-resolved fluorescence microscopy. In another push, an increasing number of funding and research institutions commit to FAIR principles as well as promoting open-science initiatives. 

The design of Luminosa`s software makes all data easily accessible. It works with the open, well documented PTU data format, enabling custom analysis. Moreover, it includes various data export options.