Crisafi Francesco, Kumar Vikas, Perri Antonio, Marangoni Marco, Cerullo Giulio, Polli Dario
IFN-CNR, Dipartimento di Fisica, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano, Italy.
IFN-CNR, Dipartimento di Fisica, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano, Italy; Center for Nano Science and Technology @Polimi, Istituto Italiano di Tecnologia, via Giovanni Pascoli 70/3, 20133 Milano, Italy.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Jan 5;188:135-140. doi: 10.1016/j.saa.2017.06.055. Epub 2017 Jul 5.
We present a multimodal non-linear optical (NLO) laser-scanning microscope, based on a compact fiber-format excitation laser and integrating coherent anti-Stokes Raman scattering (CARS), stimulated Raman scattering (SRS) and two-photon-excitation fluorescence (TPEF) on a single platform. We demonstrate its capabilities in simultaneously acquiring CARS and SRS images of a blend of 6-μm poly(methyl methacrylate) beads and 3-μm polystyrene beads. We then apply it to visualize cell walls and chloroplast of an unprocessed fresh leaf of Elodea aquatic plant via SRS and TPEF modalities, respectively. The presented NLO microscope, developed in house using off-the-shelf components, offers full accessibility to the optical path and ensures its easy re-configurability and flexibility.
我们展示了一种多模态非线性光学(NLO)激光扫描显微镜,它基于紧凑的光纤型激发激光器,并在单个平台上集成了相干反斯托克斯拉曼散射(CARS)、受激拉曼散射(SRS)和双光子激发荧光(TPEF)。我们展示了其同时获取6μm聚甲基丙烯酸甲酯珠和3μm聚苯乙烯珠混合物的CARS和SRS图像的能力。然后,我们分别通过SRS和TPEF模式将其应用于可视化伊乐藻未处理新鲜叶片的细胞壁和叶绿体。所展示的NLO显微镜是使用现成组件在内部开发的,提供了对光路的完全访问,并确保了其易于重新配置和灵活性。