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通过阴性选择富集小鼠脾细胞，以实现更广泛的单细胞转录组学研究。

Mouse splenocyte enrichment strategies via negative selection for broadened single-cell transcriptomics.

机构信息

Department of Biology, University of Nebraska at Omaha, Omaha, NE 68182, USA.

Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE 68106, USA.

出版信息

STAR Protoc. 2022 May 17;3(2):101402. doi: 10.1016/j.xpro.2022.101402. eCollection 2022 Jun 17.

DOI:10.1016/j.xpro.2022.101402
PMID:35600930
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9120244/
Abstract

Mammalian splenic tissue is rich in functional immune cells, primarily lymphocytes which can mask low-abundance populations in downstream analyses. This protocol enriches minority immune cell populations from mouse spleen via immunomagnetic negative depletion to generate an untouched enriched cell fraction. Enriched cells are then spiked with untouched splenocytes in a controlled repopulation, validated by flow cytometry and results in a single-cell transcriptomic clustering analysis with a broadened cellular landscape.

摘要

哺乳动物脾脏组织富含功能免疫细胞，主要是淋巴细胞，它们可以掩盖下游分析中低丰度群体。本方案通过免疫磁珠阴性耗竭从鼠脾脏中富集稀有免疫细胞群体，从而生成未经处理的富集细胞部分。然后，将富集细胞与未经处理的脾细胞进行受控再植入，通过流式细胞术进行验证，最终进行单细胞转录组聚类分析，扩大细胞景观。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/89d61464e477/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/6ba9afb3c521/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/ab00ffa9c63f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/59af4dec92c4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/aefc20868e2b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/8b3fdcdf3afd/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/2638fdcdc5fc/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/da4c52e11f7a/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/3339552f6cc6/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/ed249189c90e/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/89d61464e477/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/6ba9afb3c521/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/ab00ffa9c63f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/59af4dec92c4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/aefc20868e2b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/8b3fdcdf3afd/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/2638fdcdc5fc/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/da4c52e11f7a/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/3339552f6cc6/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/ed249189c90e/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df08/9120244/89d61464e477/gr9.jpg

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An updated guide for the perplexed: cytometry in the high-dimensional era.《困惑者最新指南：高维时代的细胞计数法》
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3
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4
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6
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