We report three-dimensional fluorescence emission difference(3D-FED)microscopy using a spatial light modulator(SLM).Zero phase,0–2vortex phase and binary 0-pi phase are loaded on the SLM to generate the corresponding solid,doughnut and z-axis hollow excitation spot,respectively.Our technique achieves super-resolved image by subtracting three di®erently acquired images with proper subtractive factors.Detailed theoretical analysis and simulation tests are proceeded to testify the performance of 3D-FED.Also,the improvement of lateral and axial resolution is demonstrated by imaging 100 nm°uorescent beads.The experiment yields lateral resolution of 140 nm and axial resolution of approximate 380 nm.
Guangyuan ZhaoZihao RongCuifang KuangCheng ZhengXu Liu
本文报道了一种基于拉锥结构的全光纤型内窥光学相干层析成像探针.基于大纤芯多模光纤的低光束发散特性,使用大纤芯多模光纤代替透镜作为成像元件,并在单模光纤与大纤芯多模光纤之间引入过渡拉锥段以减少插入损耗.首先利用光学仿真软件(Rsoft)确定探针的最佳结构,然后通过拉锥、切割以及熔接工艺实现探针制作,并对探针的出射光束特性与插入损耗进行测量,最后将该探针与扫频光学相干层析成像主系统联机,对人体指尖皮肤及鸡气管壁组织进行成像.该探针直径为250μm,不锈钢保护管外径为325μm,硬端长度1 cm,插入损耗约为0.3 d B,空气中有效成像范围达800μm.该探针为内窥光学相干层析成像技术在心血管疾病的应用提供了高紧凑度、高传输效率与高灵活性的选择.
Fourier ptychographic microscopy(FPM)is a newly developed imaging technique which stands out by virtue of its high-resolution and wide FOV.It improves a microscope's imaging perfor-mance beyond the diffraction limit of the employed optical components by illuminating the sample with oblique waves of different incident angles,similar to the concept of synthetic aperture.We propose to use an objective lens with high-NA to generate oblique illuminating waves in FPM.We demonstrate utilizing an objective lens with higher NA to iluminate the sample leads to better resolution by simulations,in which a resolution of 0.28 pum is achieved by using a high-NA illuminating objective lens(NA=1.49)and a low-NA collecting objective lens(NA=0.2)in coherent imaging(λ=488 nm).We then deeply study FPM's exact relevance of convergence speed to spatial spectrum overlap in frequency domain.The simulation results show that an overlap of about 60%is the optimal choice to acquire a high-quality recovery(520*520 pixels)with about 2 min's computing time.In addition,we testify the robustness of the algorithm of FPM to additive noises and its suitability for phase objects,which further proves FPM's potential application in biomedical imaging.
