光学学报, 2016, 36 (8): 0818001, 网络出版: 2016-08-18
基于波前相位调制的宽视场光片显微镜研究 下载: 550次
Field of View-Extended Light Sheet Microscope Based on Wavefront Phase Modulation
显微 荧光显微镜 视场扩展 波前相位调制 空间光调制器 microscopy fluorescence microscope field of view extending wavefront phase modulation spatial light modulator
摘要
为解决光片显微镜成像视场范围小的问题,通过光束波前相位调制与图像拼接技术实现了光片显微镜对样本的宽视场成像。数值模拟了照明光束在波前相位调制后物镜聚焦面处的光强分布。搭建了光片显微镜光路系统,并对荧光微球、菊花花粉进行成像实验。采用片状照明光束对不同聚焦位置处的样本进行成像,再将图像进行剪裁、拼接得到宽视场成像结果。实验结果表明,488 nm激发光通过数值孔径为0.3的物镜照明样本成像,可在保持光片厚度为0.81 μm的情况下达到31.93 μm的成像视场,视场扩展到原视场的3倍以上。仿真和实验表明,采用光束波前相位调制与图像拼接技术可在不损失层切能力的前提下扩展光片显微镜的成像视场。
Abstract
In order to extend the field of view (FOV) of the light sheet microscopy system, wavefront phase modulation and digital image tiling are proposed. The intensity distribution of the light sheet at the focal plane of the objective lens after wavefront phase modulation is numerically simulated. A light sheet microscopy system is built to carry out the imaging experiments for fluorescent particles and flower pollen. The light sheet illuminates different sample positions by beam defocusing to obtain different images, which are tiled to get the FOV-extended image. FOV is extended to 3 times (about 31.93 μm) of the original field with wavelength of 488 nm and numerical aperture of 0.3 while the thickness of light sheet retains 0.81 μm. The experiment and simulation results suggest that FOV of the system can be extended by wavefront phase modulation and digital image tiling with tomographic ability not sacrificed.
徐豪, 张运海, 张欣, 黄维. 基于波前相位调制的宽视场光片显微镜研究[J]. 光学学报, 2016, 36(8): 0818001. Xu Hao, Zhang Yunhai, Zhang Xin, Huang Wei. Field of View-Extended Light Sheet Microscope Based on Wavefront Phase Modulation[J]. Acta Optica Sinica, 2016, 36(8): 0818001.