Department of Physics, University of Osnabrück, Barbarastr. 7, 49069 Osnabrück, Germany.
J Biophotonics. 2013 Oct;6(10):785-92. doi: 10.1002/jbio.201200148. Epub 2012 Dec 7.
Immuno-SERS microscopy is a novel imaging technique in nano-biophotonics, which employs antibodies labeled with SERS-active nanoparticles in conjunction with Raman microscopy. Rapid data acquisition is of central importance for screening large areas of tissue specimens. Here, we first discuss the role of SERS labels with single-particle sensitivity in immuno-SERS microscopy, in particular with respect to false-negative results. In combined single-particle experiments (SERS microscopy/dark-field microscopy/HR-SEM), we then demonstrate that small glass-coated clusters (dimers and trimers) of gold nanospheres exhibit the desired single-particle SERS sensitivity, even at acquisition times as short as 30 msec per pixel, while monomers do not. The proof-of-concept for rapid immuno-SERS microscopy with 30 msec acquisition time per pixel for selective imaging of the p53 family member p63 in prostate tissue sections is demonstrated.
免疫 SERS 显微镜是纳米生物光子学中的一种新型成像技术，它结合了带有 SERS 活性纳米粒子的抗体和拉曼显微镜。快速的数据采集对于筛选大片组织标本至关重要。在这里，我们首先讨论了具有单粒子灵敏度的 SERS 标记物在免疫 SERS 显微镜中的作用，特别是在假阴性结果方面。然后，在结合的单粒子实验（SERS 显微镜/暗场显微镜/高分辨率扫描电子显微镜）中，我们证明了即使在每个像素 30 毫秒的极短采集时间下，小的玻璃覆盖的金纳米球（二聚体和三聚体）簇也表现出所需的单粒子 SERS 灵敏度，而单体则没有。我们证明了在前列腺组织切片中，通过 30 毫秒采集时间的快速免疫 SERS 显微镜可以选择性地对 p53 家族成员 p63 进行成像，其采集时间为每个像素 30 毫秒。
