PO.CL01.14 · 临床研究

用于多组学SpaceIQ™平台中空间解析分子成像数据的端到端质量控制流程

An end-to-end quality control pipeline for spatially resolved molecular imaging data in the multi-omic SpaceIQ™ platform

海报缩略图:用于多组学SpaceIQ™平台中空间解析分子成像数据的端到端质量控制流程
编号 6675 展板 17 时间 4/21 02:00–05:00 区域 Section 48 主讲 Brian Falkenstein, MS
分会场 Spatial Proteomics and Transcriptomics 3
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作者与单位 Authors & Affiliations

Brian Falkenstein1, A. Burak Tosun1, Raymond Yan1, S. Chakra Chennubhotla2, Filippo Pullara1

1PredxBio, Inc., Pittsburgh, PA,2PredxBio, Inc. / University of Pittsburgh, Pittsburgh, PA

摘要 Abstract

中文摘要
背景:mIF成像的快速扩展增加了对严格、标准化质量控制的需求。样本制备、染色和图像采集过程中引入的变异性可能掩盖生物学信号并损害下游分析。常见问题包括扭曲前景-背景对比的批次效应、导致过曝/欠曝或低对比度的采集误差,以及气泡、碎屑或脱蜡缺陷等物理伪影。更难检测的是非特异性抗体结合,它可能出现在意料之外的结构或细胞类型中。这些挑战需要一个人机协同(human-in-the-loop)的QC系统,能够在多样的组织和平台上检测、纠正并记录质量偏差。 方法:我们使用一个泛组织真实世界mIF数据集,评估了SpaceIQ™ QC流程在最常见的图像质量失效模式下的表现。QC从原始像素数据开始:由细胞核衍生的组织掩膜界定前景/背景区域,用于估计和纠正批次效应。通道级参数模型检测模糊、饱和、气泡和其他采集相关伪影。空间建模量化组织区域间的不均匀照明。使用预期染色模式库来识别非特异性结合。分割输出用于标记生物学上不可能的共表达事件(如PanCK/CD45),作为染色或采集问题的指标。每个QC模块生成一个0-3分的评分,从低质量/不可用到无伪影,实现简单、可解释的定量评估。所有QC输出均在SpaceIQ™界面中可视化,该界面高亮显示含伪影的ROI,并支持用户确认或覆盖。 结果:该流程可靠地检测并排除了受模糊、饱和和不均匀照明影响的区域。应用标准化QC步骤通过纠正批次效应和移除受非特异性染色影响的区域,显著改变了细胞计数和亚型分布,证明了在定量分析前进行系统性QC的重要性。 结论:SpaceIQ™ QC流程为mIF成像的自动化而可解释的质量评估提供了一个不依赖于平台和组织的框架。通过整合伪影检测、批次纠正和生物学有效性检查,它提高了准确性、可重复性和用户信心,确保结果反映真实的生物学信号而非技术性变异。
查看英文原文 English abstract
Background: The rapid expansion of mIF imaging has increased the need for rigorous, standardized quality control. Variability introduced during sample prep, staining, and image acquisition can obscure biological signals and compromise downstream analysis. Common issues include batch effects that distort foreground-background contrast, acquisition errors causing over/underexposure or low contrast, and physical artifacts such as bubbles, debris, or deparaffinization defects. More difficult to detect is non-specific antibody binding, which can appear in unexpected structures or cell types. These challenges require a human-in-the-loop QC system capable of detecting, correcting, and documenting quality deviations across diverse tissues and platforms. Methods: Using a pan-tissue real-world mIF dataset, we evaluated the SpaceIQ™ QC pipeline across the most frequent image quality failure modes. QC begins with raw pixel data: a nuclear-derived tissue mask defines foreground/background regions for estimating and correcting batch effects. Channel-level parametric models detect blurring, saturation, bubbles, and other acquisition-related artifacts. Spatial modeling quantifies uneven illumination across tissue areas. A library of expected staining patterns is used to identify non-specific binding. Segmentation outputs are used to flag biologically impossible co-expression events (e.g., PanCK/CD45) as indicators of staining or acquisition issues. Each QC module generates a 0-3 score, from low quality/not-usable to no-artifacts, enabling a simple, interpretable quantitative assessment. All QC outputs are visualized in the SpaceIQ™ interface, which highlights artifact-containing ROIs and enables user confirmation or override. Results: The pipeline reliably detected and excluded regions affected by blurring, saturation, and uneven illumination. Applying standardized QC steps significantly altered cell counts and subtype distributions by correcting batch effects and removing regions affected by non-specific staining, demonstrating the importance of systematic QC before quantitative analysis. Conclusions: The SpaceIQ™ QC pipeline provides a platform- and tissue-agnostic framework for automated yet interpretable quality assessment of mIF imaging. By integrating artifact detection, batch correction, and biological validity checks, it improves accuracy, reproducibility, and user confidence, ensuring results reflect true biological signal rather than technical variability.
利益披露 Disclosure
B. Falkenstein, None.. A. Tosun, None.. R. Yan, None.. S. Chennubhotla, None.. F. Pullara, None.

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