Characterization of autofluorescence and quantitative protoporphyrin IX biomarkers for optical spectroscopy-guided glioma surgery
Open accessAbstract
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 PpIX 620 contribution, thus halving the mean Ratio 620/634 ( p < 0.001). The ratio was approximately 0 for HGGs and increasing for LGGs, as demonstrated previously. Additionally, the Ratio 620/634 , 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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Most high-grade gliomas show strong PpIX fluorescence, whereas low-grade gliomas typically do not exhibit PpIX fluorescence.
“Although most high-grade gliomas show strong PpIX fluorescence, low-grade gliomas typically do not exhibit PpIX fluorescence [50,51].”
From 5-Aminolevulinic Acid as a Theranostic Agent for Tumor Fluorescence Imaging and Photodynamic Therapy (2023) · cites this paper as [50]
The Introduction (page unavailable) says only about 20% of low-grade gliomas display *visible* PpIX fluorescence during surgery, and the Results report that fluorescence visibility generally increases with malignancy. But the paper does not establish that most high-grade gliomas show *strong* fluorescence; it also says spectroscopy can detect PpIX when a tumor is not visibly fluorescing. The claim therefore omits an important distinction between no visible fluorescence and no PpIX fluorescence. (LLM, conf=0.82)
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Knab, A., Marupally, A. G., Anwer, A. G., Habibalahi, A., Pattamatta, U., Carnt, N. A., Casson, R. J., Goldys, E. M., & White, A. J. R. (2026) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2026.09.017
Goff, N. K., Markert, J. E., Park, M., Springer, A., Chen, A. M., & Orringer, D. A. (2026) · Neurosurgical FOCUS · doi:10.3171/2026.8.focus26439
- Authors
- Black, D., Kaneko, S., Walke, A., König, S., Stummer, W., & Suero Molina, E.
- Affiliations
- Carl Zeiss (Germany) (DE)
- Hokkaido University (JP)
- University of Münster (DE)
- University Hospital Münster (DE)
- Venue
- Scientific Reports
- Published
- 2021-10-08
- Volume
- 11
- Issue
- 1
- Pages
- 20009-20009
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC8501114
- OpenAlex
- W3202000232
- Cited by
- 46
- References
- 42
- Citations by year
- 2026: 7 · 2025: 16 · 2024: 15 · 2023: 6 · 2022: 2
- FWCI
- 3.04
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- URN:NBN Resolver for Germany & Switzerland (German National Library) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Universitäts- und Landesbibliothek Münster (cc-by)
- Funders
- Westfälische Wilhelms-Universität Münster
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
- Health Sciences › Medicine › Genetics › Glioma Diagnosis and Treatment
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Photodynamic Therapy Research Studies
- Keywords
- glioma surgery, 5-aminolevulinic acid, protoporphyrin IX, autofluorescence, hyperspectral fluorescence imaging, spectral unmixing, low-grade glioma, high-grade glioma, tumor infiltration, NADH, lipofuscin, flavin, WHO grade
- MeSH terms
- Blood-Brain Barrier, Brain Neoplasms, Fluorescent Dyes, Glioma, Humans, Neuroglia, Protoporphyrins, Biomarkers, Tumor, Surgery, Computer-Assisted, Optical Imaging
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