光学微操纵过程的轴平面显微成像技术

光学俘获技术利用光与物质相互作用产生的光势阱效应来实现对微粒的操控,已经成功应用于生物医学、材料科学等交叉领域.在对微粒进行三维俘获时,传统的宽场光学显微技术只能观测到某一平面内微粒的横向运动,对微粒沿轴向运动的观测受到很大限制.本文将轴平面显微成像技术引入光学微粒操控研究中,利用45?倾斜的反射镜把微粒的轴向运动信息转换到横向平面进行观测,与传统宽场显微成像技术相结合,实现了对二氧化硅小球俘获过程横向和轴向运动的同步观测.该成像方法无需扫描和数据重构,具有实时快速等优点,在新型光束光镊、厚样品三维观测和成像等领域具有潜在的应用价值.

Observation of particle manipulation with axial plane optical microscopy

Optical micromanipulation of particles based on the optical trapping effect induced by the interaction between light and particles has been successfully applied to many interdisciplinary fields including biomedicine and material sciences. When particles are trapped in three dimensions, the conventional wide-field optical microscopy can only monitor the movement of the trapped particles in a certain transverse plane. The ability to observe the particle movement along light trajectories is limited. Recently, a novel method named axial plane optical microscopy(APOM) has been developed to directly image the axial plane that is parallel to the optical axis of an objective lens. The APOM observes the axial plane by converting the axial information of a sample into that of a transverse plane by using a 45°-tilted mirror. In this paper, we propose and demonstrate that the APOM serves as an effective tool for observing the axial movement of particles in optical tweezers. By combining with a conventional wide-field optical microscopy, we show that both transverse and axial information can be acquired simultaneously for the optical micromanipulation. As in our first experimental demonstration, we observe two particles which are trapped and aligned along the optical axis. From the transverse image, only one particle is observable, and it is difficult to obtain the information along the axial direction. However, in the axial plane imaging, the longitudinal dipolar structure formed by the two particles is clearly visible. This clearly demonstrates the APOM imaging capability along the axial axis. The numerically simulations on the trapping focal spot against the position of a collimating lens agree well with our experimental APOM results. Furthermore, we directly observe the dynamic capture process of a single trapped particle in transverse plane by conventional wide-field optical microscopy as well in axial plane by the APOM, and can obtain the 3D information rapidly and simultaneously. We point out that the observable axial dynamic range is about 30 μm. Taking advantages of no requirement of scanning and data reconstruction, the APOM has potential applications in many fields, including optical trapping with novel beams and 3D imaging of thick biological specimens.