The fluorescent tracer 5-aminolevulinc acid was introduced to visualize brain tumors intraoperatively, but suffers from drawbacks such as limited sensitivity for certain tumor types. Optical coherence tomography (OCT) is a non-invasive imaging modality, which has recently found its application in neuroscience by contributing label-free tissue information. We present one of the first radiomics-based analyses to capture the form and texture of glioma samples resected during fluorescence-guided surgery in a large cohort of multimodal OCT-based microscopy (OCM) imaging data. Concluding, we report encouraging results for the prediction of tumor infiltration, entity and molecular biomarkers with accuracies as high as 96%.
Maximal safe resection is key to prolonging overall survival during intracranial surgery. Capabilities of optical coherence tomography (OCT) to discriminate tissue types have been previously reported. We utilized an OCT-based microscope (OCM) to acquire reflectivity data of resected brain tumor samples and perform textural analyses. Twelve features showed statistically significant differences between different tissue types. Three of those – contrast, energy and entropy – are described in detail in this work. Contrast and entropy increased with tissue malignancy, and energy decreased. These results suggest that OCM data-based textural analyses are capable of differentiating different tissue types based on their intrinsic structures.
In this pilot study, a multi-modal optical coherence microscopy (OCM) and fluorescence imaging system in the visible wavelength region was utilized to investigate the intra-tumor heterogeneity of lower grade brain tumor samples. OCM intensity data derived attenuation coefficients were calculated and cell density maps were created from intensity en-face projections. Sequentially preoperatively administrated 5-ALA fluorescence intensities were evaluated using the multi-modal setup. Optical imaging data results were compared to histology-derived cell density and cell proliferation rate maps.
Protoporphyrin IX (PpIX) fluorescence-guided surgery has established as a standard for resecting malign glioma. However, low-grade glioma or sparsely infiltrated brain often emit weak PpIX fluorescence and are hard to distinguish from non-pathological tissue. Furthermore, spectrally overlapping autofluorescence inherently limits the sensitivity of fluorescence-intensity based PpIX detection. We therefore integrated frequency-domain fluorescence lifetime imaging together with a spectrometer in a surgical microscope. When analyzing human glioma samples ex vivo, weak PpIX fluorescence could be differentiated from the autofluorescence background through increased lifetimes. Characteristic peaks in the spectral measurements (635, 705nm) confirmed low concentrations of PpIX in the tissue.
Maximal safe resection is of utmost importance when operating on intracranial tumors. However, the discrimination between physiological and malignant tissue is challenging. A solution is 5-aminolevulinic acid (5-ALA) induced protoporphyrin IX (PPIX) fluorescence of malignant cells, which has evolved as powerful intraoperative tool for tumor identification. We investigated 43 samples obtained during 5-ALA guided surgery using a combined optical coherence microscopy (OCM) and fluorescence imaging system and compared results to cell density maps utilizing digital pathology. Based on fluorescence intensity, attenuation coefficients, and cell density, tumor core was differentiated from tumor-associated brain parenchyma with high sensitivity and specificity.
We utilized a visible light optical coherence microscopy imaging (OCM) system to investigate the intra-tumor heterogeneity of glioblastoma samples. Using a broad visible spectrum an axial resolution of 0.8 μm was achieved. Objective lenses with short focal lengths were integrated to achieve high transverse resolution. Large field of view images were acquired using an automatic moving x-y-stage. Attenuation maps were calculated from OCM data and cell-density evaluations were performed from corresponding histological sections. Cell density and attenuation data were evaluated. Our analysis revealed intra-tumor heterogeneity by varying cell densities and attenuation coefficients throughout the tumorous areas.
Significance: 5-Aminolevulinic acid (5-ALA)-based fluorescence guidance in conventional neurosurgical microscopes is limited to strongly fluorescent tumor tissue. Therefore, more sensitive, intrasurgical 5-ALA fluorescence visualization is needed.
Aim: Macroscopic fluorescence lifetime imaging (FLIM) was performed ex vivo on 5-ALA-labeled human glioma tissue through a surgical microscope to evaluate its feasibility and to compare it to fluorescence intensity imaging.
Approach: Frequency-domain FLIM was integrated into a surgical microscope, which enabled parallel wide-field white-light and fluorescence imaging. We first characterized our system and performed imaging of two samples of suspected low-grade glioma, which were compared to histopathology.
Results: Our imaging system enabled macroscopic FLIM of a 6.5 × 6.5 mm2 field of view at spatial resolutions <20 μm. A frame of 512 × 512 pixels with a lifetime accuracy <1 ns was obtained in 65 s. Compared to conventional fluorescence imaging, FLIM considerably highlighted areas with weak 5-ALA fluorescence, which was in good agreement with histopathology.
Conclusions: Integration of macroscopic FLIM into a surgical microscope is feasible and a promising method for improved tumor delineation.
5-aminolevulinic acid positive brain tumor samples were investigated with a multimodal visible light optical coherence microscope (OCM) and fluorescence imaging (FI) setup. Tumor specific contrast was achieved by the FI part while the three-dimensional tissue morphology was assessed by the OCM part of the setup.
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