Vertical data collected by Scanning Electron Microscopy (SEM) are important for sample characterization, 3D reconstruction, and flex manipulation. Traditional methods are limited by the extent to which the probe obstructs the view of the sample along the vertical axis. Herein, we propose a novel SEM microprobe for measuring the vertical distance between the probe and substrate. To form a semi-transparent hole that is set as the objective regions in processing of the SEM images, an epoxy film was embedded in the through-hole at the tip of the microforce probe with 3D printing. The film can be modified with a focused ion beam (FIB) system. The motion of the modified probe along the vertical axis is controlled by a nanopositioner and the process is recorded by taking a real-time SEM video. The change in gray contrast caused by the semi-transparent epoxy is corrected during the SEM image processing of the video. By comparing the gray contrast with the nanopositioner motion data, we find that the change in gray contrast can provide feedback for adjusting the displacement between the probe and the substrate, and the resolution can be up to 100 nm. We propose a novel and simple method for measuring vertical distances in the SEM, which is useful for in-situ measurements and nanomanipulations.

通过扫描电子显微镜（SEM）收集的垂直数据对于样品表征，3D重建和弯曲操作非常重要。传统方法受到探针沿垂直轴遮挡样品视线的程度的限制。本文中，我们提出了一种新颖的SEM微探针，用于测量探针和基板之间的垂直距离。为了形成在SEM图像处理中设置为目标区域的半透明孔，将环氧树脂膜通过3D打印嵌入到微力探针尖端的通孔中。可以使用聚焦离子束（FIB）系统对薄膜进行改性。修饰探针沿垂直轴的运动由纳米定位器控制，并通过拍摄实时SEM视频记录该过程。在视频的SEM图像处理过程中，校正了由半透明环氧树脂引起的灰色对比度变化。通过将灰色对比度与纳米定位器运动数据进行比较，我们发现灰色对比度的变化可以提供反馈，以调整探针和基板之间的位移，并且分辨率可以达到100 nm。我们提出了一种新颖而简单的方法来测量SEM中的垂直距离，这对于原位测量和纳米操作很有用。