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基于双轴锥镜的多层面快速自动对焦系统设计

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针对现有显微成像系统对透明样品对焦时存在抗干扰性差、易脱焦、光能利用率低等问题,计算、设计并搭建了一种基于双轴锥镜的半环形光束自动对焦系统。为了获得环形光束内、外径的计算公式,利用斯涅尔定律与几何关系,获得了环形光束口径与轴锥镜折射率、间距之间的相互关系;由于系统光源的非对称性,为了计算系统的空间分辨率和轴向分辨率,采用分割近似和振幅叠加原理,推导了半环形光束的远场振幅分布,可计算系统分辨能力;为了验证光束质量并与实验相互验证,根据菲涅耳-基尔霍夫理论推导了高斯光束经过轴锥镜后的准确光能分布公式。设计并搭建了工作波长为785 nm、半环形光束内/外径为19 mm/20 mm、成像焦距为200 mm的快速自动对焦系统,其中成像光路在400~790 nm范围实现了宽谱段消色差,避免了不同波长对焦不一致;照明光路获得了边界清晰锐利的环形光束光能分布,为多层面识别提供了良好照明条件;在物镜数值孔径(NA)为0。8时,计算得到对焦系统的空间分辨率为0。77 μm,轴向分辨率为0。40 µm,小于显微物镜景深的1/4,满足高精度对焦要求。
Design of a Multilayer Fast Autofocus System Based on Dual-Axis Conical Mirrors
Objective Autofocus technology significantly enhances the imaging process,enabling rapid target acquisition and improving productivity and efficiency.It plays a crucial role in modern technological development.Currently,autofocus technologies can be categorized into three primary methods:the pre-scan focus map method,the deep learning calculation method,and the real-time reflective focus method.Each has its advantages and limitations.For example,the pre-scan focus map method requires multiple images and offers poor anti-interference capabilities;the deep learning calculation method has a limited focusing range and requires extensive system training;the real-time reflective focus method,although the most widely used,presents several challenges.Achieving real-time,fast,multi-level autofocus with a wide focusing range remains an urgent issue in the industry.Methods In this paper,we propose a multilayer automatic focusing method based on biaxial axicon mirrors.A collimated laser beam passes through two inverted axicon mirrors with matching angles,resulting in a collimated,parallel annular beam.After being blocked,a semi-annular beam is produced,which propagates through the system.Upon reaching the sample plane via a beam splitter and objective lens,multiple surfaces on the object side reflect the semi-annular beam to the objective lens.The returned beam is then reflected by the beam splitter and imaged onto a CCD or CMOS sensor.Using the conjugate relationship between the object and image,multiple semi-annular spots are formed on the image plane,sharing a common center but without overlapping.The energy center position of each spot correlates with the defocus of the sample,enabling the determination of the defocus amount and direction of different reflective surfaces.Through closed-loop control of a servo motor,fast automatic focusing across multiple levels can be achieved.Results and Discussions Through simulation and experimental setups,a multilayer autofocus system based on a Nikon 20× objective lens is designed,as shown in Figs.6 and 10.Using Eq.(4),the axial resolution of the system is calculated to be 0.4 μm,as shown in Fig.8.A comparison between Zemax software simulations and experimental results confirms the theoretical accuracy.By analyzing Eq.(11),the result shown in Fig.9(a)aligns with Tracepro software simulations,demonstrating that the beam edge obtained with this method is clear and easy to interpret.The system's achromatic performance and defocusing simulations(Fig.7)meet the expected outcomes.The experimental setup of the multilayer autofocus system,as shown in Fig.11(a),is consistent with both theoretical and simulation results.Conclusions In this paper,we propose a novel multilayer focusing method based on axicon mirrors,highlighting its advantages in terms of diffraction effects,energy efficiency,and beam selection.We derive the relationship between the aperture and width of the circular beam,the refractive index of the axicon,and the distance between axicon mirrors,providing direct experimental guidance.A method for aperture segmentation and energy superposition is proposed to calculate the resolution of the semi-annular beam.Using Fresnel-Kirchhoff diffraction theory,the specific energy distribution of Gaussian beams after passing through an axicon mirror is derived.Based on this theory,an automatic focusing system using a 20× objective lens is designed and constructed.The annular beam's aperture is 20 mm,the axicon refractive index is 1.4585,and the distance between the biaxial axicon mirrors is 123 mm.The system achieves a spatial resolution of 0.77 μm and an axial resolution of 0.40 μm,and the focusing accuracy is about 1/4 depth of field of the microscope objective lens.The theory's validity is verified through both simulations and experiments,demonstrating that the beam quality produced by this method is ideal for applications requiring multilayer autofocus,such as in biomedical and electronic circuit fields.

autofocusaxicon mirrormultilayeroptical designmicroscopy

李林晗、刘英、于晨、周广鹏、王成龙、党博石、马俊林、杜杰

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中国科学院长春光学精密机械与物理研究所精密仪器与装备研发中心,吉林 长春 130033

中国科学院大学,北京 101408

自动对焦 轴锥镜 多层面 光学设计 显微镜

2024

光学学报
中国光学学会 中国科学院上海光学精密机械研究所

光学学报

CSTPCD北大核心
影响因子:1.931
ISSN:0253-2239
年,卷(期):2024.44(23)