PO.CH02.02 · 化学
由ZEISS slide stream和Vizgen InSituPlex®检测实现的用于空间生物学的高通量、自动化、可重现mIF工作流程
A high-throughput, automated, and reproducible mIF workflow for spatial biology enabled by ZEISS slide stream and Vizgen's InSituPlex® assays
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摘要 Abstract
中文摘要
多重免疫荧光（mIF）已成为空间生物学中研究肿瘤微环境的基石技术。尽管应用广泛，现有的mIF工作流程往往受到图像采集变异性以及对劳动密集型手动步骤依赖的制约。为了克服这些局限，我们开发了一套完全整合的高通量空间生物学工作流程，将高度标准化和自动化的ZEISS空间生物学平台与Vizgen预优化的InSituPlex检测相结合。二者共同实现了在宽动态范围内高通量、完全自动化和可重现的生物标志物检测，同时提高了效率和可扩展性。为验证这些说法，我们开展了一项多中心验证研究。使用包含CD3、Ki67、Granzyme-B和CK/Sox10的4重OmniVUE面板对多种组织适应症的福尔马林固定石蜡包埋（FFPE）切片进行染色。在所有中心，切片均采用相同的预定义自动化流程，使用Leica Bond RX仪器染色，并使用ZEISS Axioscan 7空间生物学切片扫描仪成像。采集的图像无缝上传至Mindpeak，并使用针对在Axioscan扫描仪上成像的Vizgen InSituPlex检测优化的集成STARVUE算法进行分析，提供阳性细胞密度和荧光强度的定量分析。结果分析表明各中心之间具有出色的一致性，证实了该组合工作流程在多个操作员和仪器间的精密度和准确度。定性方面，各中心的染色高度可比，荧光信号和背景的有限变异并未影响STARVUE™ AI算法的准确性。定量方面，所展示的工作流程在不同地点间表现出高重现性和可重复性，为转化研究提供了可靠的解决方案，并提升了数据生成的速度和质量。因此，它为推动空间生物学从转化研究走向临床应用提供了重大潜力。
查看英文原文 English abstract
Multiplex immunofluorescence (mIF) has become a cornerstone technique in spatial biology for investigating the tumor microenvironment. Despite its widespread use, existing mIF workflows are often hindered by variability in image acquisition and dependence on labor-intensive manual steps. To overcome these limitations, we developed a fully integrated, high-throughput spatial biology workflow combining the highly standardized and automated ZEISS spatial biology platform with Vizgen's pre-optimized InSituPlex assay. Together, these deliver high-throughput, fully automated, and reproducible biomarker detection across a broad dynamic range, enhancing both efficiency and scalability. To validate these claims, we conducted a multisite verification study. A 4-plex OmniVUE panel including CD3, Ki67, Granzyme-B and CK/Sox10 was utilized to stain multiple tissue indications on formalin-fixed paraffin-embedded (FFPE) sections. On all sites, the slides underwent the same predefined and automated routine for staining using Leica Bond RX instruments and imaging using ZEISS Axioscan 7 spatial biology slide scanners. Acquired images were seamlessly uploaded to Mindpeak and analyzed with integrated STARVUE algorithms optimized for Vizgen InSituPlex assays imaged on Axioscan scanners, providing quantitative analysis for densities and fluorescence intensities of positive cells. Analysis of the results demonstrated excellent concordance between sites, confirming the precision and accuracy of the combined workflow across multiple operators and instruments. Qualitatively, staining was highly comparable across all sites, and the limited variations in fluorescence signal and background did not affect the accuracy of the STARVUE™ AI algorithms. Quantitatively, the presented workflow exhibits high reproducibility and repeatability of results across locations, providing a reliable solution for translational research with amplified speed and quality of data generation. As a result, it offers significant potential to advance spatial biology from translational research to clinical applications
利益披露 Disclosure
C. Kysilovsky, None..
K. Hwang, None..
S. Pfeifenbring, None..
M. Downing, None..
S. Derakhshani, None..
M. Widmaier, None..
J. He, None..
A. Vasaturo, None.