Modern optics offers several label-free microscopic and spectroscopic solutions which are useful for both imaging and pathological assessments of biological tissues. The possibility to obtain similar morphological and biochemical information with fast and label-free techniques is highly desirable, but no single optical modality is capable of obtaining all of the information provided by histological and immunohistochemical analyses. Integrated multimodal imaging offers the possibility of integrating morphological with functional-chemical information in a label-free modality, complementing the simple observation with multiple specific contrast mechanisms. Here, we developed a custom laser-scanning microscopic platform that combines confocal Raman spectroscopy with multimodal non-linear imaging, including Coherent Anti-Stokes Raman Scattering, Second-Harmonic Generation, Two-Photon Excited Fluorescence, and Fluorescence Lifetime Imaging Microscopy. The experimental apparatus is capable of high-resolution morphological imaging of the specimen, while also providing specific information about molecular organization, functional behavior, and molecular fingerprint. The system was successfully tested in the analysis of ex vivo tissues affected by urothelial carcinoma and by atherosclerosis, allowing us to multimodally characterize of the investigated specimen. Our results show a proof-of-principle demonstrating the potential of the presented multimodal approach, which could serve in a wide range of biological and biomedical applications.
Understanding the deterioration processes in wooden artefacts is essential for accurately assessing their conservation status and developing effective preservation strategies. Advanced imaging techniques are currently being explored to study the impact of chemical changes on the structural and mechanical properties of wood. Nonlinear optical modalities, including second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), combined with fluorescence lifetime imaging microscopy (FLIM), offer a promising non-destructive diagnostic method for evaluating lignocellulose-based materials. In this study, we employed a nonlinear multimodal approach to examine the effects of artificially induced delignification on samples of Norway spruce (Picea abies) and European beech (Fagus sylvatica) subjected to increasing treatment durations. The integration of SHG/TPEF imaging and multi-component fluorescence lifetime analysis enabled the detection of localized variations in nonlinear signals and τ-phase of key biopolymers within wood cell walls. This methodology provides a powerful tool for early detection of wood deterioration, facilitating proactive conservation efforts of wooden artefacts.
Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults, accounting for more than 60% of all cases.1 Current treatment options at diagnosis include surgical resection, radiation, and chemotherapy2 but—despite advances in this field—it remains largely incurable; its 5-year survival rate3 is . Extensive and complete surgical resection of GBM is difficult due to the high invasiveness of these tumors. However, multiple studies4,5 have demonstrated the importance of aggressive surgical resection—when possible—with trends toward better outcomes in those patients with a greater extent of resection.