FLUORESCENT DYES IN MODERN NEUROSURGERY: TECHNOLOGICAL ADVANCES AND CLINICAL TRANSLATION
DOI:
https://doi.org/10.31435/ijitss.2(50).2026.5241Keywords:
Fluorescence-Guided Surgery, 5-Aminolevulinic Acid, Fluorescein, Indocyanine Green, Neurosurgery, Intraoperative ImagingAbstract
Introduction and purpose: Over recent decades, neurosurgery has undergone major technological advancement toward increased precision and safety, including the development of fluorescence-guided surgery. Conventional white-light microscopy has limited ability to distinguish pathological from normal tissue or to assess vascular flow in real time. Fluorescent dyes enhance intraoperative visualization and support surgical decision-making. This review aims to summarize the properties, mechanisms of action, clinical applications, and safety profiles of fluorescent dyes used in contemporary neurosurgery.
Materials and methods: A comprehensive literature search was conducted in PubMed, Scopus, Web of Science, and Google Scholar (2000–2025) using keywords including fluorescence-guided surgery, fluorescent dyes, neurosurgery, 5-aminolevulinic acid, fluorescein, indocyanine green, ICG angiography, glioma surgery, brain tumor resection, aneurysm clipping, vascular neurosurgery, near-infrared imaging, intraoperative imaging, and optical navigation. Eligible studies were qualitatively analyzed and synthesized narratively.
Conclusion: Fluorescent dyes have become integral tools in modern neurosurgery, enhancing visualization of tumors and vascular structures beyond conventional white-light microscopy. 5-ALA, fluorescein, and indocyanine green each provide unique mechanisms of fluorescence, supporting tailored intraoperative decision-making. Overall, these agents are well-tolerated, with severe adverse events being rare. Future advancements are likely to focus on integrating fluorescence with machine learning, augmented reality, quantitative assessment, and novel imaging technologies, improving precision and objectivity. While their current clinical position is stable, ongoing technological evolution may expand applications and refine existing techniques, maintaining fluorescence-guided surgery as a cornerstone of precision neurosurgical practice.
References
Leone, A., Carbone, F., Spetzger, U., Vajkoczy, P., Raffa, G., Angileri, F., Germanó, A., Engelhardt, M., Picht, T., Colamaria, A., & Rosenstock, T. (2025). Preoperative mapping techniques for brain tumor surgery: A systematic review. Frontiers in Oncology, 14, Article 1481430. https://doi.org/10.3389/fonc.2024.1481430
Carbone, M., Montemurro, N., Cattari, N., Autelitano, M., Cutolo, F., Ferrari, V., Cigna, E., & Condino, S. (2025). Targeting accuracy of neuronavigation: A comparative evaluation of an innovative wearable AR platform vs. traditional EM navigation. Frontiers in Digital Health, 6, Article 1500677. https://doi.org/10.3389/fdgth.2024.1500677
Katal, S., York, B., & Gholamrezanezhad, A. (2024). AI in radiology: From promise to practice—A guide to effective integration. European Journal of Radiology, 181, Article 111798.
Najjar, R. (2023). Redefining radiology: A review of artificial intelligence integration in medical imaging. Diagnostics, 13(17), Article 2760. https://doi.org/10.3390/diagnostics13172760
Kantarcı, M., Aydın, S., Oğul, H., & Kızılgöz, V. (2025). New imaging techniques and trends in radiology. Diagnostic and Interventional Radiology, 31(5), 505–517. https://doi.org/10.4274/dir.2024.242926
Drummen, G. P. (2012). Fluorescent probes and fluorescence (microscopy) techniques—Illuminating biological and biomedical research. Molecules, 17(12), 14067–14090. https://doi.org/10.3390/molecules171214067
Hickey, S. M., Ung, B., Bader, C., Brooks, R., Lazniewska, J., Johnson, I. R. D., Sorvina, A., Logan, J., Martini, C., Moore, C. R., Karageorgos, L., Sweetman, M. J., & Brooks, D. A. (2021). Fluorescence microscopy—An outline of hardware, biological handling, and fluorophore considerations. Cells, 11(1), Article 35. https://doi.org/10.3390/cells11010035
Sanderson, M. J., Smith, I., Parker, I., & Bootman, M. D. (2014). Fluorescence microscopy. Cold Spring Harbor Protocols, 2014(10), pdb.top071795. https://doi.org/10.1101/pdb.top071795
Luu, P., Fraser, S. E., & Schneider, F. (2024). More than double the fun with two-photon excitation microscopy. Communications Biology, 7, Article 364.
Marshall, J., & Johnsen, S. (2017). Fluorescence as a means of colour signal enhancement. Philosophical Transactions of the Royal Society B: Biological Sciences, 372(1724), Article 20160335. https://doi.org/10.1098/rstb.2016.0335
Dunst, S., & Tomancak, P. (2019). Imaging flies by fluorescence microscopy: Principles, technologies, and applications. Genetics, 211(1), 15–34. https://doi.org/10.1534/genetics.118.300227
Datta, R., Heaster, T. M., Sharick, J. T., Gillette, A. A., & Skala, M. C. (2020). Fluorescence lifetime imaging microscopy: Fundamentals and advances in instrumentation, analysis, and applications. Journal of Biomedical Optics, 25(7), 1–43. https://doi.org/10.1117/1.JBO.25.7.071203
Vicidomini, G., Bianchini, P., & Diaspro, A. (2018). STED super-resolved microscopy. Nature Methods, 15, 173–182.
Filip, P., Lerner, D. K., Kominsky, E., Schupper, A., Liu, K., Khan, N. M., Roof, S., Hadjipanayis, C., Genden, E., & Iloreta, A. M. C. (2024). 5-Aminolevulinic acid fluorescence-guided surgery in head and neck squamous cell carcinoma. Laryngoscope, 134(2), 741–748. https://doi.org/10.1002/lary.30910
Hadjipanayis, C. G., Widhalm, G., & Stummer, W. (2015). What is the surgical benefit of utilizing 5-aminolevulinic acid for fluorescence-guided surgery of malignant gliomas? Neurosurgery, 77(5), 663–673. https://doi.org/10.1227/NEU.0000000000000929
Liu, Z., Mela, A., Argenziano, M. G., Banu, M. A., Furnari, J., Kotidis, C., Sperring, C. P., Humala, N., Mahajan, A., Bruce, J. N., Canoll, P., & Sims, P. A. (2023). Single-cell analysis of 5-aminolevulinic acid intraoperative labeling specificity for glioblastoma. Journal of Neurosurgery, 140(4), 968–978. https://doi.org/10.3171/2023.7.JNS23122
Schupper, A. J., Baron, R. B., Cheung, W., Rodriguez, J., Kalkanis, S. N., Chohan, M. O., Andersen, B. J., Chamoun, R., Nahed, B. V., Zacharia, B. E., Kennedy, J., Moulding, H. D., Zucker, L., Chicoine, M. R., Olson, J. J., Jensen, R. L., Sherman, J. H., Zhang, X., Price, G., Fowkes, M., Germano, I. M., Carter, B. S., Hadjipanayis, C. G., & Yong, R. L. (2021). 5-Aminolevulinic acid for enhanced surgical visualization of high-grade gliomas: A prospective, multicenter study. Journal of Neurosurgery, 136(6), 1525–1534. https://doi.org/10.3171/2021.5.JNS21310
Iinuma, S., Bachor, R., Flotte, T., & Hasan, T. (1995). Biodistribution and phototoxicity of 5-aminolevulinic acid-induced PpIX in an orthotopic rat bladder tumor model. Journal of Urology, 153(3 Pt 1), 802–806.
Yahanda, A. T., Dunn, G. P., & Chicoine, M. R. (2021). Photosensitivity reaction from operating room lights after oral administration of 5-aminolevulinic acid for fluorescence-guided resection of a malignant glioma. Cureus, 13(2), Article e13442. https://doi.org/10.7759/cureus.13442
Wei, L., Roberts, D. W., Sanai, N., & Liu, J. T. C. (2019). Visualization technologies for 5-ALA-based fluorescence-guided surgeries. Journal of Neuro-Oncology, 141(3), 495–505. https://doi.org/10.1007/s11060-018-03077-9
Valerio, J. E., de Jesús Aguirre Vera, G., Zumaeta, J., Rea, N. S., Fernandez Gomez, M. P., Mantilla-Farfan, P., Valente, L., & Alvarez-Pinzon, A. M. (2025). Comparative analysis of 5-ALA and fluorescent techniques in high-grade glioma treatment. Biomedicines, 13(5), Article 1161. https://doi.org/10.3390/biomedicines13051161
Jin, E., Yin, H., Gui, Y., Chen, J., Zhang, J., Liang, J., Li, X. X., & Zhao, M. (2019). Fluorescein angiographic findings of peripheral retinal vasculature after intravitreal conbercept versus ranibizumab for retinopathy of prematurity. Journal of Ophthalmology, 2019, Article 3935945. https://doi.org/10.1155/2019/3935945
Kakucs, C., Florian, I. A., Ungureanu, G., & Florian, I. S. (2017). Fluorescein angiography in intracranial aneurysm surgery: A helpful method to evaluate the security of clipping and observe blood flow. World Neurosurgery, 105, 406–411. https://doi.org/10.1016/j.wneu.2017.05.172
Cavallo, C., De Laurentis, C., Vetrano, I. G., Falco, J., Broggi, M., Schiariti, M., Ferroli, P., & Acerbi, F. (2018). The utilization of fluorescein in brain tumor surgery: A systematic review. Journal of Neurosurgical Sciences, 62(6), 690–703. https://doi.org/10.23736/S0390-5616.18.04480-6
Wolfe, D. R. (1986). Fluorescein angiography basic science and engineering. Ophthalmology, 93(12), 1617–1620. https://doi.org/10.1016/s0161-6420(86)33521-8
Diaz, R. J., Dios, R. R., Hattab, E. M., Burrell, K., Rakopoulos, P., Sabha, N., Hawkins, C., Zadeh, G., Rutka, J. T., & Cohen-Gadol, A. A. (2015). Study of the biodistribution of fluorescein in glioma-infiltrated mouse brain and histopathological correlation of intraoperative findings in high-grade gliomas resected under fluorescein fluorescence guidance. Journal of Neurosurgery, 122(6), 1360–1369. https://doi.org/10.3171/2015.2.JNS132507
Kofoed, M. S., Pedersen, C. B., Schulz, M. K., Kristensen, B. W., Hansen, R. W., Markovic, L., Halle, B., & Poulsen, F. R. (2022). Fluorescein-guided resection of cerebral metastases is associated with greater tumor resection. Acta Neurochirurgica, 164(2), 451–457. https://doi.org/10.1007/s00701-021-04796-1
Morykwas, M. J., Hills, H., & Argenta, L. C. (1991). The safety of intravenous fluorescein administration. Annals of Plastic Surgery, 26(6), 551–553. https://doi.org/10.1097/00000637-199106000-00009
Restelli, F., Bonomo, G., Monti, E., Broggi, G., Acerbi, F., & Broggi, M. (2022). Safeness of sodium fluorescein administration in neurosurgery: Case-report of an erroneous very high-dose administration and review of the literature. Brain & Spine, 2, Article 101703. https://doi.org/10.1016/j.bas.2022.101703
Danis, R. P., & Stephens, T. (1997). Phototoxic reactions caused by sodium fluorescein. American Journal of Ophthalmology, 123(5), 694–696.
Šranková, M., Dvořák, A., Martínek, M., Šebej, P., Klán, P., Vítek, L., & Muchová, L. (2022). Antiproliferative and cytotoxic activities of fluorescein—a diagnostic angiography dye. International Journal of Molecular Sciences, 23(3), Article 1504. https://doi.org/10.3390/ijms23031504
Dzurinko, V. L., Gurwood, A. S., & Price, J. R. (2004). Intravenous and indocyanine green angiography. Optometry, 75(12), 743–755. https://doi.org/10.1016/s1529-1839(04)70234-1
Desmettre, T., Devoisselle, J. M., & Mordon, S. (2000). Fluorescence properties and metabolic features of indocyanine green (ICG) as related to angiography. Survey of Ophthalmology, 45(1), 15–27.
Pollmann, L., Juratli, M., Roushansarai, N., Pascher, A., & Hölzen, J. P. (2023). Quantification of indocyanine green fluorescence imaging in general, visceral and transplant surgery. Journal of Clinical Medicine, 12(10), Article 3550. https://doi.org/10.3390/jcm12103550
Nguyen, A. M., Do, H. H., Huynh Trung, N., & Bui, D. H. T. (2025). Preliminary results of paraclinoid aneurysm clipping with indocyanine green–video angiography: A single-center experience in Vietnam. World Neurosurgery, 197, Article 123928.
Avery, M., Chehab, S., Wong, J. H., & Mitha, A. P. (2016). Intraoperative indocyanine green videoangiography to guide decision making regarding need for vessel bypass: A case report and technical note. Surgical Neurology International, 7(Suppl 2), S36–S39. https://doi.org/10.4103/2152-7806.173567
Catapano, G., Sgulò, F., Laleva, L., Columbano, L., Dallan, I., & de Notaris, M. (2018). Multimodal use of indocyanine green endoscopy in neurosurgery: A single-center experience and review of the literature. Neurosurgical Review, 41(4), 985–998. https://doi.org/10.1007/s10143-017-0858-4
Hope-Ross, M., Yannuzzi, L. A., Gragoudas, E. S., Guyer, D. R., Slakter, J. S., Sorenson, J. A., Krupsky, S., Orlock, D. A., & Puliafito, C. A. (1994). Adverse reactions due to indocyanine green. Ophthalmology, 101(3), 529–533. https://doi.org/10.1016/s0161-6420(94)31303-0
Prünte, C., & Flammer, J. (1996). Choroidal capillary and venous congestion in central serous chorioretinopathy. American Journal of Ophthalmology, 121(1), 26–34. https://doi.org/10.1016/s0002-9394(14)70531-8
Kruiswijk, M. W., Nguyen Dinh, H. L., Tange, F. P., Koning, S., van den Hoven, P., Peul, R. C., Rotmans, J. I., Huurman, V. A. L., Alwayn, I. P. J., Hamming, J. F., Vahrmeijer, A. L., & van der Vorst, J. R. (2024). The safety of indocyanine green in patients with advanced chronic kidney disease or kidney transplantation: A scoping review. Annals of Medicine and Surgery, 87(3), 1351–1359. https://doi.org/10.1097/MS9.0000000000002713
Afridi, A., Zulfiqar, A., Sajjad, F., Shahid, I., Habib, H., Saleem, Y., Afridi, Z., Iqbal, A., Khattak, F., Nisa, F., Khan, H., Bacha, Z., Ali, M. A., Khan, M. H., Afridi, R., & Kamil, K. A. (2025). Efficacy and safety of indocyanine green fluorescence imaging in colorectal cancer: A systematic review and meta-analysis of randomized controlled trials. International Journal of Colorectal Disease, 40(1), Article 193. https://doi.org/10.1007/s00384-025-04941-7
Li, J., Lan, Z., He, M., & You, C. (2009). Assessment of microscope-integrated indocyanine green angiography during intracranial aneurysm surgery: A retrospective study of 120 patients. Neurology India, 57(4), 453–459. https://doi.org/10.4103/0028-3886.55607
Doss, V. T., Goyal, N., Humphries, W., Hoit, D., Arthur, A., & Elijovich, L. (2015). Comparison of intraoperative indocyanine green angiography and digital subtraction angiography for clipping of intracranial aneurysms. Interventional Neurology, 3(3–4), 129–134. https://doi.org/10.1159/000381148
Chen, J. S., Young, J. S., & Berger, M. S. (2025). Current and future applications of 5-aminolevulinic acid in neurosurgical oncology. Cancers, 17(8), Article 1332. https://doi.org/10.3390/cancers17081332
Mousa, A. H., Scalia, G., Ahmed, N., Figueiredo, E. G., de Sena Barbosa, M. G., Khan, M. M., Chavez-Herrera, V. R., & Chaurasia, B. (2025). The evolution of intracranial aneurysm clipping: A historical perspective. Annals of Medicine and Surgery, 87(4), 2229–2233. https://doi.org/10.1097/MS9.0000000000003162
Rodriguez, B., Brown, C. S., Colan, J. A., Zhang, J. Y., Huq, S., Rivera, D., Young, T., Williams, T., Subramaniam, V., & Hadjipanayis, C. (2025). Fluorescence-guided surgery for gliomas: Past, present, and future. Cancers, 17(11), Article 1837. https://doi.org/10.3390/cancers17111837
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Jędrzej Sztajura, Karol Józef Szkarłat, Maksymilian Szklarski , Szymon Targosz, Michał Stachel, Ewa Maraszewska, Weronika Szymacha , Aleksandra Płecka, Karol Zimnicki, Alicja Stępień, Katarzyna Nycz, Olha Levchenko, Rafał Kobylański

This work is licensed under a Creative Commons Attribution 4.0 International License.
All articles are published in open-access and licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0). Hence, authors retain copyright to the content of the articles.
CC BY 4.0 License allows content to be copied, adapted, displayed, distributed, re-published or otherwise re-used for any purpose including for adaptation and commercial use provided the content is attributed.

