胶质母细胞瘤是一种侵袭性很强的肿瘤，是最常见的原发性脑恶性肿瘤。主要特征包括其对放疗和化疗等常规治疗的高抵抗力及其弥漫性组织浸润，阻止完全手术切除。对生理微环境中的迁移和侵袭过程的分析可以增强对这些现象的理解，并可以改进治疗方法。在这里，我们将两种最先进的技术结合起来，即成人器官型脑组织切片培养 (OTC) 和透明组织的光片荧光显微镜 (LSFM)，称为 OTCxLSFM 的组合方法。利用这种方法，我们可以证明，通过三氧化二砷或 WP1066 治疗，以及四跨膜蛋白、跨膜受体 CD9 或信号转导子和转录激活子 3 (STAT3) 的基因缺失，可以有效阻止胶质母细胞瘤组织浸润。通过我们的分析流程，我们获得了单细胞水平的三维信息，以及对肿瘤细胞形态外观的深入了解。© 2023 作者。根据 CC BY NC ND 4.0 许可证条款发布。

Glioblastoma is a very aggressive tumor and represents the most common primary brain malignancy. Key characteristics include its high resistance against conventional treatments, such as radio- and chemotherapy and its diffuse tissue infiltration, preventing complete surgical resection. The analysis of migration and invasion processes in a physiological microenvironment allows for enhanced understanding of these phenomena and can lead to improved therapeutic approaches. Here, we combine two state-of-the-art techniques, adult organotypic brain tissue slice culture (OTC) and light-sheet fluorescence microscopy (LSFM) of cleared tissues in a combined method termed OTCxLSFM. Using this methodology, we can show that glioblastoma tissue infiltration can be effectively blocked through treatment with arsenic trioxide or WP1066, as well as genetic depletion of the tetraspanin, transmembrane receptor CD9, or signal transducer and activator of transcription 3 (STAT3). With our analysis pipeline, we gain single-cell level, three-dimensional information, as well as insights into the morphological appearance of the tumor cells.© 2023 The Authors. Published under the terms of the CC BY NC ND 4.0 license.