Raman nanoparticle probes for antibody-based protein detection in tissues
Raman nanoparticle probes for antibody-based protein detection in tissues
复制标题
DOI：
10.1369/jhc.7a7313.2007
复制
发表时间：
2008-04-01
影响因子：
3.2
通讯作者：
Knudsen, Beatrice
中科院分区：
文献类型：
--
作者：
Lutz, Barry;Dentinger, Claire;Knudsen, Beatrice
表面增强拉曼散射（SERS）纳米粒子正在成为生物分子光学检测的新方法。在福尔马林固定石蜡包埋 (FFPE) 前列腺组织切片的模型测定中，我们使用与复合有机-无机纳米粒子 (COIN) 缀合的抗体 (Ab) 检测前列腺特异性抗原 (PSA)，并使用相同的染色方案来比较相邻组织切片中的 COIN-Ab 和 Alexa-Ab 缀合物。光谱分析阐明了荧光和拉曼特征之间的根本区别，并从背景自发荧光中准确提取 COIN 探针信号。探针信号用于生成组织上 PSA 表达的图像，并提供质量测量来表征 COIN 检测与 Alexa 相比的性能。 COIN 的染色准确性（正确识别上皮细胞中 PSA 表达的能力）略低于 Alexa，这是由于 COIN 的假阴性率较高。然而，COIN 提供了与 Alexa 相当的信号强度，以及良好的批次内、批次间和批次间一致性。总体而言，COIN 和 Alexa 检测试剂与 FFPE 组织具有相似的性能，支持进一步开发用于该应用的拉曼探针。本手稿包含 http://www.jhc.org 上的在线补充材料。请在线访问本文以查看这些材料。
Surface-enhanced Raman scattering (SERS) nanoparticles are emerging as a new approach for optical detection of biomolecules. In a model assay in formalin-fixed paraffin-embedded (FFPE) prostate tissue sections, we detect prostate-specific antigen (PSA) using antibody (Ab) conjugated to composite organic-inorganic nanoparticles (COINs), and we use identical staining protocols to compare COIN-Ab and Alexa-Ab conjugates in adjacent tissue sections. Spectral analysis illustrates the fundamental difference between fluorescence and Raman signatures and accurately extracts COIN probe signals from background autofluorescence. Probe signals are used to generate images of PSA expression on the tissue, and quality measures are presented to characterize the performance of the COIN assay in comparison to Alexa. Staining accuracy (ability to correctly identify PSA expression in epithelial cells) is somewhat less for COIN than Alexa, which is attributed to an elevated false negative rate of the COIN. However, COIN provided signal intensities comparable to Alexa, and good intra-, inter-, and lot-to-lot consistencies. Overall, COIN and Alexa detection reagents possess similar performance with FFPE tissues, supporting the further development of Raman probes for this application. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.