Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, United States.
J Phys Chem B. 2011 May 12;115(18):5574-81. doi: 10.1021/jp1113834. Epub 2011 Apr 19.
Label-free microscopy based on Raman scattering has been increasingly used in biomedical research to image samples that cannot be labeled or stained. Stimulated Raman scattering (SRS) microscopy allows signal amplification of the weak Raman signal for fast imaging speeds without introducing the nonresonant background and coherent image artifacts that are present in coherent anti-Stokes Raman scattering (CARS) microscopy. Here we present the Raman-induced Kerr effect (RIKE) as a contrast for label-free microscopy. RIKE allows us to measure different elements of the nonlinear susceptibility tensor, both the real and imaginary parts, by optical heterodyne detection (OHD-RIKE). OHD-RIKE microscopy provides information similar to polarization CARS (P-CARS) and interferometric CARS (I-CARS) microscopy, with a simple modification of the two-beam SRS microscopy setup. We show that, while OHD-RIKE microspectroscopy can be in principle more sensitive than SRS, it does not supersede SRS microscopy of heterogeneous biological samples, such as mouse skin tissue, because it is complicated by variations of linear birefringence across the sample.
基于拉曼散射的无标记显微镜在生物医学研究中越来越多地被用于对无法标记或染色的样本进行成像。受激拉曼散射(SRS)显微镜允许对弱拉曼信号进行信号放大,从而实现快速成像速度,而不会引入在相干反斯托克斯拉曼散射(CARS)显微镜中存在的非共振背景和相干图像伪影。在这里,我们提出拉曼诱导克尔效应(RIKE)作为无标记显微镜的对比度。RIKE 允许我们通过光学外差检测(OHD-RIKE)测量非线性磁化率张量的不同元素,包括实部和虚部。OHD-RIKE 显微镜通过对双光束 SRS 显微镜设置进行简单修改,提供类似于偏振 CARS(P-CARS)和干涉 CARS(I-CARS)显微镜的信息。我们表明,虽然 OHD-RIKE 微光谱学在原理上可以比 SRS 更灵敏,但它并没有取代对异质生物样本(如小鼠皮肤组织)的 SRS 显微镜,因为它受到样品中线性双折射变化的影响。