A cost- and time-efficient method for high-throughput cryoprocessing and tissue analysis using multiplexed tissue molds.

IF 4.3 Q1 BIOCHEMICAL RESEARCH METHODS

Cell Reports Methods Pub Date : 2025-04-21 DOI:10.1016/j.crmeth.2025.101023

Daniel Reumann, Martin Colombini, Paul Möseneder, Agnieszka Piszczek, Jürgen A Knoblich

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Abstract

Cryosectioning remains the gold standard for antibody and transcriptomic/in situ hybridization tissue analysis. However, tissue processing is time-consuming and costly, limiting routine and diagnostic use. Currently, no commercially available protocols or products exist for multiplexing this process. Here, we introduce multiplexed tissue molds (MTMs) that enable high-throughput cryoprocessing-cutting costs and workload by up to 96% while permitting the processing of tissues of various sizes and origins. We demonstrate compatibility with heterogeneous tissues by processing 19 different adult mouse tissues in parallel. Furthermore, we process up to ∼110 neural organoids of different ages and sizes simultaneously and assess their neural differentiation marker expression. MTMs allow sectioning-based tissue analysis when labor, time, and cost are limiting factors. MTMs could be used to compare high specimen numbers in histopathological settings, organism-wide antigen and antibody targeting studies, high-throughput tissue screens, and defined tissue section positioning for, e.g., spatial transcriptomics experiments.

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使用多路组织模具进行高通量冷冻处理和组织分析的成本和时间效率方法。

冷冻切片仍然是抗体和转录组/原位杂交组织分析的金标准。然而，组织处理既耗时又昂贵，限制了常规和诊断的使用。目前，没有商业上可用的协议或产品存在用于多路复用这个过程。在这里，我们介绍了多路组织模具（mtm），它可以实现高通量冷冻处理，在允许加工各种尺寸和来源的组织的同时，削减高达96%的成本和工作量。我们通过平行处理19种不同的成年小鼠组织来证明与异质组织的相容性。此外，我们同时处理了多达110个不同年龄和大小的神经类器官，并评估了它们的神经分化标志物表达。当人工、时间和成本是限制因素时，mtm允许基于切片的组织分析。mtm可用于比较组织病理学设置中的高标本数量，全生物体抗原和抗体靶向研究，高通量组织筛选和确定的组织切片定位，例如空间转录组学实验。

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