Recent Advances in the Analysis of Single Cells.
Recent Advances in the Analysis of Single Cells.
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DOI：
10.1021/acs.analchem.6b04255
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发表时间：
2017-01-03
影响因子：
7.4
通讯作者：
Dittrich PS
中科院分区：
文献类型：
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作者：
Armbrecht L;Dittrich PS
分析方法的进步和新兴的微流控工具使详细研究活体中的生物过程成为可能，并达到了足以进行单细胞分析的灵敏度。单细胞分析这一术语通常是指阐明大细胞群体中细胞间的差异，如大小、形态、生长速度或分子含量，如脂类、蛋白质、代谢物、DNA/RNA等的组成。已开发了许多不同的技术来解决细胞异质性的影响。1−4就我们今天所知，异质性出现在所有细胞群(细菌、酵母和哺乳动物细胞)中，甚至在细胞谱系中，所有细胞都来自完全相同的母细胞。除了基本的研究问题(如为什么细胞不同，这种差异如何影响细胞的生理和命运？)，单细胞分析在许多研究领域都有实际应用。5如本综述所述，这些例子包括癌症生物学、干细胞和再生医学、微生物学和发病机制、神经科学、免疫学等等。单细胞分析的最大挑战来自于细胞的小尺寸、目标分子的微小绝对数量、存在于广泛浓度范围内的大量不同分子以及许多相关的细胞内或细胞间动力学过程带来的复杂性。为了在单细胞水平上跟踪这些动态过程，由于对环境变化或药物、细胞分化或代谢变化的响应，除了高灵敏度和特异度外，还需要具有高时间分辨率和高通量的方法。具有高精度和准确读出的量化对于确保所揭示的异质性确实源于细胞群体而不是有条不紊的人工制品是至关重要的。到目前为止，各种化学和物理技术被应用于单细胞分析领域。它们通常涉及单个细胞的选定方面，并且可以相互补充。接下来，我们将重点介绍近两年来在荧光显微镜、电化学分析、质谱学和定量聚合酶链式反应技术等领域的新进展。由于微流控方法被应用于
Advances of analytical methods and emerging microfluidic tools have made it possible to investigate biological processes in living organisms in detail and reach sensitivities sufficient for single-cell analysis. The term “single-cell analysis” typically refers to the elucidation of cell-to-cell differences in large cell populations, such as size, morphology, growth rate, or molecular content like the composition of lipids, proteins, metabolites, DNA/RNA, etc. Many different techniques have been developed to address the effects of cell heterogeneities. 1− 4 As far as we know today, heterogeneities appear in all cell populations (bacteria, yeast, and mammalian cells) and even within cell lineages, where all cells are derived from the very same mother cell. Besides the fundamental research questions (such as, why are cells different and how does the difference affect cell physiology and fate?), single-cell analysis has practical applications in many research fields. 5 As will be covered in this Review, the examples include cancer biology, stem cells and regenerative medicine, microbiology and pathogenesis, neuroscience, immunology, and many more. The biggest challenges of single-cell analysis arise from the small size of cells, the tiny absolute number of target molecules, the large number of different molecules present in a wide range of concentrations and, last but not least, the complexity imposed by many related intra-or intercellular dynamic processes. To follow these dynamic processes at the single cell level, due to the response to environmental changes or drugs, cell differentiation, or metabolic changes, methods with a high time resolution and high throughput are required in addition to high sensitivity and specificity. Quantification with highly precise and accurate read-out is essential to ensure that the revealed heterogeneities indeed originate from the cell population and are not methodical artifacts. To date, various chemical and physical techniques are applied in the field of single-cell analysis. They typically address selected aspects of the single cells and may be complementary to each other. In the following, we focus on new developments in the fields of fluorescence microscopy, electrochemical analysis, mass spectrometry, and qPCR based technologies in the last two years. As microfluidic methods are employed in
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影响因子：
16.6
作者：
Burns JC;Kelly MC;Hoa M;Morell RJ;Kelley MW
通讯作者：
Kelley MW
影响因子：
16.8
作者：
Bendall SC;Nolan GP;Roederer M;Chattopadhyay PK
通讯作者：
Chattopadhyay PK
DOI：
10.1126/science.1198704
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Bendall SC;Simonds EF;Qiu P;Amir el-AD;Krutzik PO;Finck R;Bruggner RV;Melamed R;Trejo A;Ornatsky OI;Balderas RS;Plevritis SK;Sachs K;Pe'er D;Tanner SD;Nolan GP
通讯作者：
Nolan GP
影响因子：
4.6
作者：
Brinkmann F;Hirtz M;Haller A;Gorges TM;Vellekoop MJ;Riethdorf S;Müller V;Pantel K;Fuchs H
通讯作者：
Fuchs H
影响因子：
82.9
作者：
通讯作者：
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