Analysis of the Porphyrin Peak Shift and Fluorescence Lifetime in Gliomas with Different Tumor Grades, Intratumoral Regions, and Visible Fluorescence Status

Background: 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) fluorescence shows high sensitivity in detecting the tumor core of high-grade gliomas (HGG) but poor sensitivity for tissue of low-grade gliomas (LGG) and the margins of HGG. The characteristic emission peak for PpIX is known to be located at 635 nm. Recently, a second emission peak was described at 620 nm wavelength in LGG and the tumor infiltration zone of HGG. Methods: During surgery, samples from the tumor core and tumor infiltration zone of 43 WHO grade 2-4 gliomas were collected after preoperative 5-ALA administration, and their PpIX emission spectra, as well as fluorescence lifetimes, were determined by ex vivo analysis. Subsequently, the relative PpIX peak contribution (RPPC) was retrieved by calculating the integral of the two bands corresponding to the two emission peaks of PpIX (615-625 nm, 625-635 nm) and correlated with fluorescence lifetimes. Results: The mean RPPC decreased in samples with descending order of WHO grades, non-fluorescing samples, and infiltrative tumor regions, indicating a shift toward the 620 nm peak in porphyrin fluorescence. The porphyrin peak shift across all specimens correlated with lower fluorescence lifetimes (R: 0.854, R-squared: 0.729). Conclusions: The observed peak shift has important implications for fluorescence lifetime analyses since the lifetimes of other porphyrins contribute to the overall decay dynamics. Based on these initial data using fluorescence lifetime, this knowledge is of major importance, especially for detecting tissue from LGG that lack visible fluorescence, to further optimize the visualization of these tumor tissue using this promising imaging modality.