Our new Technical Note demonstrates the possibility of identifying distinct metabolic profiles of organoids using Fluorescence Lifetime Imaging (FLIM) of autofluorescence arising from different bound and unbound states of NAD(H) under two-photon excitation.
Fluorescence lifetime images were acquired using the Luminosa single-photon counting laser scanning confocal microscope from PicoQuant. Luminosa combines intuitive workflows, online image previews for real-time quality assessment, and advanced, fast GPU-based analysis routines. The fully motorized microscope system enables quick auto-alignment and power calibration for streamlined FLIM operation.
The images were further analyzed using NovaFLIM software, which allows seamless application of various analysis methods, including phasor plots, FastLifetime contrast, multi-exponential fitting, and pattern-matching analysis.