PicoQuant`s new confocal microscope Luminosa combines state-of-the-art hardware with cutting edge software to deliver high quality data while simplifying daily operation.
Luminosa’s rapidFLIM hardware can record several frames per second, 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.
We demonstrate the combination of PicoQuant’s latest multi-channel TCSPC device and a cooled high-performance 23-pixel SPAD-array developed jointly with Pi Imaging Technologies for time-resolved confocal image scanning microscopy (FLIM-ISM). We also discuss how advanced data processing can be applied to FLIM-ISM for additional performance gains.
For certain applications widefield imaging offers unique advantages. Here we show how the combination of a powerful PicoQuant laser and the novel SPAD512S camera from Pi Imaging Technology facilitates time-resolved widefield imaging. We benchmark the camera’s performance, demonstrating video rate acquisition speeds with minimal photobleaching.
Quantitative time-resolved fluorescence techniques like FLIM are increasingly employed in fields like phase separation and cellular sensing. PicoQuant`s new microscope Luminosa combines state-of-the-art hardware with cutting edge software, delivering high quality data while simplifying daily operation. The software includes features like context-based workflows, sample-free auto-alignment and laser power calibration which improve reproducibility of experiments. We describe how FLIM is streamlined with Luminosa. Its rapidFLIM hardware records several frames per second with high count rates, which the software handles with a novel dynamic binning format. Combined with GPU-accelerated algorithms, this enables high-speed automated analysis of FLIM images with minimal user interaction. We will also show an outlook about how Luminosa can be used for combining FLIM with super-resolution modalities.
Second harmonic generation (SHG) imaging microscopy is a nonlinear optical imaging technique that uses SHG as a contrast mechanism to produce high-resolution images. SHG occurs in materials with non-centrosymmetric crystal structures. Therefore, SHG imaging has been applied for characterization of 2D semiconductors, transition-metal dichalcogenides (TMDs) such as WS2 and MoSe2, lithium niobate crystals, PZT thin films, graphene, lanthanides, and even biological tissues. It provides information on the crystal lattice, assesses crystal quality and maps grain boundaries, defects, and mechanical strain. Furthermore, SHG imaging reveals the number of stacked layers as well as their orientation with respect to each other.
Fluorescence Lifetime Imaging (FLIM) has become more attractive in recent years as it offers increased specificity in many assays as well as the possibility of multiplexing the read out of many markers with a small number of detectors.
Here we present how FLIM modalities are implemented in Luminosa, the new single-photon counting confocal microscope by PicoQuant. Thanks to a dynamic binning format and GPU-based algorithms FLIM images of 1024x1024 can be analysed in a few seconds. The FLIM analysis workflow suggests the best fitting model based on statistical arguments and requires minimal user interaction making these modalities become accessible to new users who can then confidently start working with FLIM and incorporate it into their research toolbox combining the strengths of phasor plots with decay fitting.
Luminosa, the new single photon counting confocal microscope from PicoQuant addresses the challenges of an expanding single-molecule FRET community.
In this talk we present how easily these measurements can be performed with Luminosa single photon-counting confocal microscope and how all necessary correction parameters are automatically determined requiring no interaction from the user by employing methodologies benchmarked by the scientific community. We will also show how the variable PSF feature can be used in such measurements to fine-tune the observation window of freely diffusing biomolecules.
Fluorescence Lifetime Imaging (FLIM) is an essential tool in Life Sciences, but up to now users had to chose between high timing precision or fast data acquisition when using Time-Correlated Single Photon Counting (TCSPC) electronics. Our approach, named rapidFLIMHiRes, allows recording several FLIM images per second with a temporal resolution of 10 ps. The method combines advances in fast scanning, hybrid photomultiplier detectors, TCSPC modules, and correction algorithms to reduce decay curve distortions. Thus fast processes can be observed with the high optical and temporal resolution achievable in confocal microscopy at a rate of several frames per second.
Fluorescence Lifetime Imaging (FLIM) is an essential tool in Life Sciences, but up to now users had to chose between high timing precision or fast data acquisition when using Time-Correlated Single Photon Counting (TCSPC) electronics. Our approach, named rapidFLIMHiRes, allows recording several FLIM images per second with a temporal resolution of 10 ps. The method combines advances in fast scanning, hybrid photomultiplier detectors, TCSPC modules, and correction algorithms to reduce decay curve distortions. Thus fast processes can be observed with the high optical and temporal resolution achievable in confocal microscopy at a rate of several frames per second.
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