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使用BD Rhapsody单细胞分析对复杂人体组织中的低mRNA含量细胞进行分析。

Profiling low-mRNA content cells in complex human tissues using BD Rhapsody single-cell analysis.

作者信息

Scheiber Alexandra, Trebo Manuel, Pittl Annabella, Heidegger Isabel, Hautz Theresa, Oberhuber Rupert, Trajanoski Zlatko, Augustin Florian, Sopper Sieghart, Wolf Dominik, Pircher Andreas, Salcher Stefan

机构信息

Department of Internal Medicine V, Hematology and Oncology and Comprehensive Cancer Center Innsbruck (CCCI), Medical University of Innsbruck, 6020 Innsbruck, Austria.

Department of Internal Medicine V, Hematology and Oncology and Comprehensive Cancer Center Innsbruck (CCCI), Medical University of Innsbruck, 6020 Innsbruck, Austria; Tyrolean Cancer Research Center Innsbruck, 6020 Innsbruck, Austria.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103475. doi: 10.1016/j.xpro.2024.103475. Epub 2024 Dec 9.

DOI:10.1016/j.xpro.2024.103475
PMID:39661509
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11683236/
Abstract

The successful recovery of immune cells, particularly those with low mRNA content, by single-cell RNA sequencing (scRNA-seq) remains a significant challenge. Tissue dissociation and selection of the appropriate scRNA-seq technology are crucial. Our protocol efficiently recovers low-mRNA content immune cells using the BD Rhapsody scRNA-seq platform. It includes optimized tissue dissociation for prostate, lung, and liver tissues, cell labeling with Sample Tag antibodies, microwell-based single-cell capture, cDNA synthesis, library preparation, and data pre-processing with basic quality control analysis. For complete details on the use and execution of this protocol, please refer to Salcher et al..

摘要

通过单细胞RNA测序（scRNA-seq）成功回收免疫细胞，尤其是那些mRNA含量低的细胞，仍然是一项重大挑战。组织解离和选择合适的scRNA-seq技术至关重要。我们的方案使用BD Rhapsody scRNA-seq平台有效地回收了低mRNA含量的免疫细胞。它包括针对前列腺、肺和肝组织的优化组织解离、用样品标签抗体进行细胞标记、基于微孔的单细胞捕获、cDNA合成、文库制备以及进行基本质量控制分析的数据预处理。有关此方案的使用和执行的完整详细信息，请参考Salcher等人的研究。

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