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Three-dimensional random-access confocal microscopy with 3D remote focusing system

  • Haoyang Li,
  • Quan Lu,
  • Zhong Wang,
  • Wenbo Zhang,
  • Yu Wu,
  • Yandong Sun,
  • Yue Hu,
  • Lehui Xiao,
  • Dongping Zhong,
  • Suhui Deng,
  • Shangguo Hou

摘要

Understanding biological activities in cells or deep tissues requires high-speed three-dimensional (3D) imaging. Substantial progress has been made with the emergence of 3D random-access microscopy. However, current solutions for fast 3D random-access imaging remain complex and costly. Herein we propose a simple, cost-effective, and fast 3D random-access confocal microscopy with remote focusing system. Our system shows isotropic response times across the x, y, and z axes, with a 34-fold improvement in axial response time over traditional piezo stages. We demonstrate its volumetric imaging performance with fluorescent particles and live cells. Furthermore, we validate the 3D random-access imaging capability of this system by continuously monitoring the signals in three different planes, showing a refresh rate of 500 Hz on two different positions in 3D. The simplicity, versatility, and affordability of our system promise widespread applications in research and industry.