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Abstract
5-Aminolevulinic acid (5-ALA)-mediated fluorescence does not effectively depict low grade gliomas (LGG) or the infiltrative tumor portion of high-grade gliomas (HGG). While spectroscopy improves sensitivity and precision, this is currently limited by autofluorescence and a second protoporphyrin IX (PpIX) fluorescence state at 620 nm. We investigated the autofluorescence to better characterize the present spectra and thus increase PpIX quantification precision and sensitivity. This study included 128 patients undergoing surgery for malignant glioma. 5-ALA (Gliolan) was administered before anesthesia, and fluorescence was measured using a hyperspectral device. It was found that all 2692 measured spectra consisted of contributions from 620 to 634 nm PpIX, NADH, lipofuscin, and flavins. The basis spectra were characterized and their use in spectral unmixing led to 82.4% lower fitting error for weakly fluorescing areas (p < 0.001), and 92.3% fewer false positive tumor identifications in control measurements (p = 0.0065) compared to previous works. They also decreased the PpIX620 contribution, thus halving the mean Ratio620/634 (p < 0.001). The ratio was approximately 0 for HGGs and increasing for LGGs, as demonstrated previously. Additionally, the Ratio620/634, the MIB-1/Ki-67 proliferation index, and the PpIX peak blue-shift were found to be significantly related to WHO grade, fluorescence visibility, and PpIX contribution (p < 0.001), and the value of these three as quantitative biomarkers is discussed.
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1 Carl Zeiss Meditec AG, Oberkochen, Germany (GRID:grid.424549.a) (ISNI:0000 0004 0379 7801); University of British Columbia, Vancouver, Canada (GRID:grid.17091.3e) (ISNI:0000 0001 2288 9830)
2 University Hospital of Münster, Department of Neurosurgery, Münster, Germany (GRID:grid.16149.3b) (ISNI:0000 0004 0551 4246); Hokkaido University Graduate School of Medicine, Department of Neurosurgery, Sapporo, Japan (GRID:grid.39158.36) (ISNI:0000 0001 2173 7691)
3 University Hospital of Münster, Department of Neurosurgery, Münster, Germany (GRID:grid.16149.3b) (ISNI:0000 0004 0551 4246); University of Münster, Core Unit Proteomics, Interdisciplinary Center for Clinical Research, Münster, Germany (GRID:grid.5949.1) (ISNI:0000 0001 2172 9288)
4 University of Münster, Core Unit Proteomics, Interdisciplinary Center for Clinical Research, Münster, Germany (GRID:grid.5949.1) (ISNI:0000 0001 2172 9288)
5 University Hospital of Münster, Department of Neurosurgery, Münster, Germany (GRID:grid.16149.3b) (ISNI:0000 0004 0551 4246)