基于非对称光束的焦面漂移测量和主动校正

Drift Measurement and Correction of Optical Focus Planes Based on Nonsymmetric Beam

电动化、智能化的光学显微成像系统需要能够实时测量系统的焦面漂移并进行校正,从而实现对活细胞的长时间观测和全片的病理切片扫描。设计一种探测非对称光束界面反射后的光斑位置从而进行焦面漂移测量的方法。利用ZEMAX软件光学仿真了反射光斑在不同离焦情况下的光斑形状,搭建了集成化的漂移测量模块并在商用正置显微镜上对其进行了验证。结果表明,针对60×浸油物镜,所提系统的漂移测量精度达250 nm,漂移校正的响应时间小于500 ms,满足了高分辨率长时间成像的要求。

Automated and intelligent optical microscopy necessitates the measurement and correction of the focal plane drift for many biomedical applications,such as long-term observation of living cells and pathological whole slide imaging.In this study,we designed a method to perform focal plane drift measurements based on the detection of the reflected spot position of a nonsymmetric beam.Through simulations using ZEMAX software,the shapes of the reflected spots were simulated on the sample surface at different defocus positions.Furthermore,an integrated focus plane drift measurement module was constructed and tested on a commercial upright microscope.The results reveal that the proposed module exhibits a drift measurement precision of 250 nm and a response time of less than 500 ms for a 60×oil immersion objective,satisfying the demand for long-term and high-resolution imaging.