Balu Mihaela, Mikami Hideharu, Hou Jue, Potma Eric O, Tromberg Bruce J
University of California, Irvine, Beckman Laser Institute, Laser Microbeam and Medical Program, Irvine, CA 92612, USA.
Current Affiliation: Department of Chemistry School of Science, University of Tokyo, Tokyo, Japan; Department of Chemistry, University of California, Irvine, CA 92697, USA.
Biomed Opt Express. 2016 Oct 3;7(11):4375-4387. doi: 10.1364/BOE.7.004375. eCollection 2016 Nov 1.
We present a multiphoton microscope designed for mesoscale imaging of human skin. The system is based on two-photon excited fluorescence and second-harmonic generation, and images areas of ~0.8x0.8 mm at speeds of 0.8 fps (800x800 pixels; 12 frame averages) for high signal-to-noise ratio, with lateral and axial resolutions of 0.5µm and 3.3µm, respectively. The main novelty of this instrument is the design of the scan head, which includes a fast galvanometric scanner, optimized relay optics, a beam expander and high NA objective lens. Computed aberrations in focus are below the Marechal criterion of 0.07λ rms for diffraction-limited performance. We demonstrate the practical utility of this microscope by imaging of wide areas in normal human skin.
我们展示了一种用于人体皮肤中尺度成像的多光子显微镜。该系统基于双光子激发荧光和二次谐波产生，以0.8帧/秒的速度（800×800像素；12次帧平均）对约0.8×0.8毫米的区域进行成像，以获得高信噪比，横向分辨率和轴向分辨率分别为0.5微米和3.3微米。该仪器的主要新颖之处在于扫描头的设计，其包括快速振镜扫描仪、优化的中继光学器件、光束扩展器和高数值孔径物镜。聚焦时计算出的像差低于用于衍射极限性能的0.07λ均方根的马雷夏尔准则。我们通过对正常人体皮肤的大面积成像来证明该显微镜的实际效用。
