High-Density Microcavity Array for Cell Detection: Single-Cell Analysis of Hematopoietic Stem Cells in Peripheral Blood Mononuclear Cells
High-Density Microcavity Array for Cell Detection: Single-Cell Analysis of Hematopoietic Stem Cells in Peripheral Blood Mononuclear Cells
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DOI：
10.1021/ac900535h
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发表时间：
2009-07-01
影响因子：
7.4
通讯作者：
Matsunaga, Tadashi
中科院分区：
文献类型：
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作者：
Hosokawa, Masahito;Arakaki, Atsushi;Matsunaga, Tadashi
从有限的生物样品中检测和分离特定细胞类型已经成为临床诊断和细胞生物学研究的主要挑战。在这里，我们报告了一个高密度的微腔阵列的靶细胞检测，其中数以千计的单细胞整齐地排列到10000微腔，高效率约90%的负载细胞。通过测量用靶向细胞表面标记物的抗体标记的细胞的荧光强度，在单细胞水平上专门鉴定细胞特异性免疫表型，并且通过该系统分析的人外周血中的造血干细胞（HSC）的纯度与通过常规流式细胞术获得的纯度相关。此外，通过使用显微操作器分离单细胞，从HSC确定干细胞标记物CD 34的基因表达。该技术已被证明是在单细胞水平上进行靶细胞检测和后续细胞分析研究的有效工具。
Detection and isolation of specific cell types from limited biological samples have become a major challenge in clinical diagnosis and cell biology research. Here, we report a high-density microcavity array for target cell detection in which thousands of single cells were neatly arrayed onto 10000 microcavities with high efficiency at approximately 90% of the loaded cells. Cell-specific immunophenotypes were exclusively identified at the single-cell level by measuring fluorescence intensities of cells labeled with antibodies targeting cell surface markers, and the purity of hematopoietic stem cells (HSCs) within human peripheral blood analyzed by this system was correlated with those obtained by conventional flow cytometry. Furthermore, gene expression of the stem cell marker, CD34, was determined from HSCs by isolating single cells using a micromanipulator. This technology has proven to be an effective tool for target cell detection and subsequent cellular analytical research at the single-cell level.