Quantitative multiplexed quantum dot immunohistochemistry
Quantitative multiplexed quantum dot immunohistochemistry
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DOI：
10.1016/j.bbrc.2008.06.127
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发表时间：
2008-09-19
影响因子：
3.1
通讯作者：
Byers, R.
中科院分区：
文献类型：
--
作者：
Sweeney, E.;Ward, T. H.;Byers, R.
量子点是光稳定的荧光半导体纳米晶体，具有宽激发和明亮、窄、对称的发射光谱。这些特点已经引起了相当大的兴趣，他们的应用在多重免疫组织化学生物标志物定量和共定位在临床样品。强大的定量允许生物标志物验证，并且由于临床样品数量有限，对多重染色的需求日益增长。大多数报道的多路量子点染色使用顺序方法，在高通量设置是费力和不切实际的。我们研究了序列多重染色的相关问题，并开发了一种利用量子点与生物素化一抗结合的方法，使三种抗体同时多重染色。CD34， Cytokeratin 18和cleaved Caspase 3在扁桃体组织中使用8小时方案进行三联体，每个定位以分离细胞区室。这证明了生物标志物测量方法的实用性，可以在单个石蜡组织切片上快速测量多个共定位的生物标志物，这对临床试验研究具有重要意义。(C) 2008爱思唯尔公司版权所有。
Quantum dots are photostable fluorescent semiconductor nanocrystals possessing wide excitation and bright narrow, symmetrical, emission spectra. These characteristics have engendered considerable interest in their application in Multiplex immunohistochemistry for biomarker quantification and co-localisation in clinical samples. Robust quantitation allows biomarker validation, and there is growing need for Multiplex staining due to limited quantity of clinical samples. Most reported multiplexed quantum dot staining used sequential methods that are laborious and impractical in a high-throughput setting. Problems associated with sequential Multiplex staining have been investigated and a method developed using QDs conjugated to biotinylated primary antibodies, enabling Simultaneous multiplex staining with three antibodies. CD34, Cytokeratin 18 and cleaved Caspase 3 were triplexed in tonsillar tissue using an 8 h Protocol, each localised to separate cellular compartments. This demonstrates utility of the method for biomarker measurement enabling rapid measurement of multiple co-localised biomarkers on single Paraffin tissue sections, of importance for clinical trial studies. (C) 2008 Elsevier Inc. All rights reserved.