用于中性粒细胞谱系细胞质谱流式分析的全骨髓制备
Preparation of Whole Bone Marrow for Mass Cytometry Analysis of Neutrophil-lineage Cells.
作者信息
Zhu Yanfang Peipei, Padgett Lindsey, Dinh Huy Q, Marcovecchio Paola, Wu Runpei, Hinz Denise, Kim Cheryl, Hedrick Catherine C
机构信息
Division of Inflammation Biology, La Jolla Institute for Immunology;
Division of Inflammation Biology, La Jolla Institute for Immunology.
出版信息
J Vis Exp. 2019 Jun 19(148). doi: 10.3791/59617.
In this article, we present a protocol that is optimized to preserve neutrophil-lineage cells in fresh BM for whole BM CyTOF analysis. We utilized a myeloid-biased 39-antibody CyTOF panel to evaluate the hematopoietic system with a focus on the neutrophil-lineage cells by using this protocol. The CyTOF result was analyzed with an open-resource dimensional reduction algorithm, viSNE, and the data was presented to demonstrate the outcome of this protocol. We have discovered new neutrophil-lineage cell populations based on this protocol. This protocol of fresh whole BM preparation may be used for 1), CyTOF analysis to discover unidentified cell populations from whole BM, 2), investigating whole BM defects for patients with blood disorders such as leukemia, 3), assisting optimization of fluorescence-activated flow cytometry protocols that utilize fresh whole BM.
在本文中，我们提出了一种经过优化的方案，用于在新鲜骨髓中保存中性粒细胞系细胞，以进行全骨髓质谱流式细胞术（CyTOF）分析。我们使用了一个偏向髓系的39抗体CyTOF panel，通过该方案来评估造血系统，重点关注中性粒细胞系细胞。使用开源的降维算法viSNE分析CyTOF结果，并展示数据以证明该方案的效果。基于此方案，我们发现了新的中性粒细胞系细胞群体。这种新鲜全骨髓制备方案可用于：1）CyTOF分析，以从全骨髓中发现未识别的细胞群体；2）研究血液疾病（如白血病）患者的全骨髓缺陷；3）辅助优化利用新鲜全骨髓的荧光激活流式细胞术方案。
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