Ex vivo autofluorescence analysis of gliomas and brain tissues of patients using macroscopic time-resolved fluorescence imaging
Daria A. Sachkova1, Marina V. Shirmanova1, Elena B. Kiseleva1, Ludmila S. Kuhnina2, Artem S. Grishin3, Ilya D. Shchechkin1, Vladislav I. Shcheslavskiy1, , Konstantin S. Yashin2
1Institute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, Nizhny Novgorod, Russia
2Department of Neurosurgery, Privolzhsky Research Medical University, Nizhny Novgorod, Russia
3Department of Pathology, Privolzhsky Research Medical University, Nizhny Novgorod, Russia
Abstract
Therapy for malignant gliomas remains ineffective, largely due to challenges in achieving complete tumor resection. The infiltrative growth of glioma cells into surrounding brain tissue complicates surgical removal and highlights the need for improved intraoperative diagnostics. Macroscopic fluorescence lifetime imaging (macro-FLIM) is a promising technique that enables real-time visualization of differences in tissues by measuring the autofluorescence decay of endogenous molecules such as nicotinamide adenine dinucleotide (phosphate) NAD(P)H.
The aim of the work was to study the autofluorescence differences between glioma types and healthy and peritumoral brain tissue of patients using macro-FLIM. The study was performed on freshly isolated brain tissue samples from patients with gliomas. Tumor samples (n=29), peritumoral white matter (38), normal white (n=15) and gray (n=11) matter were delivered to the laboratory within 60 minutes after resection on ice. Autofluorescence lifetime was visualized in the NAD(P)H channel using a confocal FLIM macroscanner (Becker&Hickl, Germany) (ex. 375 nm, reg. 435–485 nm).
Macro-FLIM analysis revealed differences in the NAD(P)H autofluorescence lifetime between healthy and tumor tissues. Tumor samples is characterized by higher τm values compared to normal white and gray matter. Notably, glioblastomas tumor samples showed elevated τm values among all glioma types (astrocytoma and oligodendroglioma), and also compared to its own PWM.
These results highlight macro-FLIM as a potential tool for intraoperative discrimination between glioma and healthy brain, as well as for identifying residual tumor infiltration. Improving surgical precision with this technique could enhance therapeutic outcomes. The study was supported by the Russian Science Foundation (project № 23-75-10068).
Speaker
Daria Alexandrovna Sachkova
Institute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, Nizhny Novgorod, Russia
Russia
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