用于处理和分析多路复用图像的协议提高了人体组织中淋巴细胞的识别和空间结构。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-07-25 DOI:10.1016/j.xpro.2025.103976

Justin A Smith, Drew T Bloss, MacKenzie D Williams, Amanda L Posgai, Todd M Brusko, Mark A Atkinson, Clive H Wasserfall, Maigan A Brusko

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引用次数: 0

摘要

在细胞表型和空间分析之前，人类淋巴组织的多路复用图像进行了广泛的预处理。在这里，我们提出了KINTSUGI（与新技术的知识集成：简化用户引导的图像处理），这是一个旨在让用户交互参与每个处理步骤以确保质量控制的协议。我们描述了参数调整和批量处理原始图像数据的步骤，包括照明校正、拼接、反卷积、3D-2D转换、配准和自荧光减法。然后我们详细介绍了分割、特征提取、表型和空间分析的程序。

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Protocol for processing and analyzing multiplexed images improves lymphatic cell identification and spatial architecture in human tissue.

Multiplexed images of human lymphatic tissue are extensively preprocessed before cell phenotyping and spatial analysis. Here, we present KINTSUGI (knowledge integration with new technologies: simplified user-guided image processing), a protocol designed to interactively engage the user in each processing step to ensure quality control. We describe steps for parameter tuning and batch processing of raw image data including illumination correction, stitching, deconvolution, 3D-2D conversion, registration, and autofluorescence subtraction. We then detail procedures for segmentation, feature extraction, phenotyping, and spatial analysis.

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