Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity.
Highly Sensitive and Multiplexed Protein Imaging With Cleavable Fluorescent Tyramide Reveals Human Neuronal Heterogeneity.
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DOI：
10.3389/fcell.2020.614624
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发表时间：
2020
影响因子：
5.5
通讯作者：
Guo J
中科院分区：
文献类型：
--
作者：
Liao R;Mondal M;Nazaroff CD;Mastroeni D;Coleman PD;Labaer J;Guo J
全面分析原位完整组织中蛋白质的能力对于我们理解健康和疾病至关重要。然而，现有的方法存在灵敏度低和样品通量有限的问题。为了解决这些问题，我们提出了一种使用可切割荧光酪酰胺和现成抗体的高灵敏度和多路复用原位蛋白分析方法。与现有方法相比，该方法提高了检测灵敏度，将成像时间缩短了1-2个数量级，并有可能在光学分辨率下检测完整组织中的数百种蛋白质。应用这种方法，我们研究了基因相同的细胞群体中的蛋白质表达异质性，并进行了蛋白质表达相关性分析以鉴定共调控蛋白。我们还分析了人类福尔马林固定石蜡包埋（FFPE）海马组织中的bb6000个神经元。通过将这些神经元根据其多路蛋白表达谱划分为不同的细胞簇，我们观察到海马的不同亚区域由来自不同簇的神经元组成。
The ability to comprehensively profile proteins in intact tissues in situ is crucial for our understanding of health and disease. However, the existing methods suffer from low sensitivity and limited sample throughput. To address these issues, here we present a highly sensitive and multiplexed in situ protein analysis approach using cleavable fluorescent tyramide and off-the-shelf antibodies. Compared with the current methods, this approach enhances the detection sensitivity and reduces the imaging time by 1–2 orders of magnitude, and can potentially detect hundreds of proteins in intact tissues at the optical resolution. Applying this approach, we studied protein expression heterogeneity in a population of genetically identical cells, and performed protein expression correlation analysis to identify co-regulated proteins. We also profiled >6,000 neurons in a human formalin-fixed paraffin-embedded (FFPE) hippocampus tissue. By partitioning these neurons into varied cell clusters based on their multiplexed protein expression profiles, we observed different sub-regions of the hippocampus consist of neurons from distinct clusters.
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影响因子：
64.5
作者：
Goltsev Y;Samusik N;Kennedy-Darling J;Bhate S;Hale M;Vazquez G;Black S;Nolan GP
通讯作者：
Nolan GP
影响因子：
3.7
作者：
Doré K;Labrecque S;Tardif C;De Koninck P
通讯作者：
De Koninck P
DOI：
10.1126/science.1198704
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
--
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
DOI：
10.1073/pnas.1507125112
发表时间：
2015-06-09
影响因子：
11.1
作者：
Darmanis S;Sloan SA;Zhang Y;Enge M;Caneda C;Shuer LM;Hayden Gephart MG;Barres BA;Quake SR
通讯作者：
Quake SR
影响因子：
30.8
作者：
Becskei, A;Kaufmann, BB;van Oudenaarden, A
通讯作者：
van Oudenaarden, A