Spatial Multiplex In Situ Tagging (MIST) Technology for Rapid, Highly Multiplexed Detection of Protein Distribution on Brain Tissue.
Spatial Multiplex In Situ Tagging (MIST) Technology for Rapid, Highly Multiplexed Detection of Protein Distribution on Brain Tissue.
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DOI：
10.1021/acs.analchem.1c04970
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发表时间：
2022-03-08
影响因子：
7.4
通讯作者：
Wang, Jun
中科院分区：
文献类型：
--
作者：
Reddy, Revanth;Yang, Liwei;Liu, Jesse;Liu, Zhuojie;Wang, Jun
生物样本的高度多重分析显著地推进了对疾病的生物学基础的理解，但是这些技术受到多重性的数量和处理速度的限制。在这里，我们提出了一种快速的多重方法定量检测蛋白质标记物的脑切片与细胞分辨率。这种空间多重原位标记（MIST）技术建立在MIST微阵列上，该微阵列包含数百万个携带条形码寡核苷酸的小微珠。使用标记有UV可切割寡核苷酸的抗体，组织切片上的蛋白质标记物的分布可以高保真地“打印”在MIST微阵列上。该技术在检测灵敏度、分辨率和信噪比方面的性能已通过检测脑细胞标志物得到充分表征。我们展示了在2小时内同时检测31种蛋白质，比其他具有类似多重性的基于免疫荧光的方法快约10倍。开发了一整套计算工具包来分割小区域并识别整个小鼠大脑的区域差异。该技术可以快速、方便地检测组织标本上的数十种生物标志物，在临床病理学和疾病机制研究中具有广泛的应用前景。
Highly multiplexed analysis of biospecimens significantly advances the understanding of biological basics of diseases, but these techniques are limited by the number of multiplexity and the speed of processing. Here, we present a rapid multiplex method for quantitative detection of protein markers on brain sections with the cellular resolution. This spatial multiplex in situ tagging (MIST) technology is built upon a MIST microarray that contains millions of small microbeads carrying barcoded oligonucleotides. Using antibodies tagged with UV cleavable oligonucleotides, the distribution of protein markers on a tissue slice could be “printed” on the MIST microarray with high fidelity. The performance of this technology in detection sensitivity, resolution and signal-to-noise level has been fully characterized by detecting brain cell markers. We showcase the codetection of 31 proteins simultaneously within 2 h which is about 10 times faster than the other immunofluorescence-based approaches of similar multiplexity. A full set of computational toolkits was developed to segment the small regions and identify the regional differences across the entire mouse brain. This technique enables us to rapidly and conveniently detect dozens of biomarkers on a tissue specimen, and it can find broad applications in clinical pathology and disease mechanistic studies.
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影响因子：
14.9
作者：
Sunkin SM;Ng L;Lau C;Dolbeare T;Gilbert TL;Thompson CL;Hawrylycz M;Dang C
通讯作者：
Dang C
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：
64.8
作者：
Harris CR;Millman KJ;van der Walt SJ;Gommers R;Virtanen P;Cournapeau D;Wieser E;Taylor J;Berg S;Smith NJ;Kern R;Picus M;Hoyer S;van Kerkwijk MH;Brett M;Haldane A;Del Río JF;Wiebe M;Peterson P;Gérard-Marchant P;Sheppard K;Reddy T;Weckesser W;Abbasi H;Gohlke C;Oliphant TE
通讯作者：
Oliphant TE
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
影响因子：
16.6
作者：
Giedt RJ;Pathania D;Carlson JCT;McFarland PJ;Del Castillo AF;Juric D;Weissleder R
通讯作者：
Weissleder R