Acta Optica Sinica, Volume. 42, Issue 14, 1411001(2022)

Three-Dimensional Multifocal Two-Photon Laser Scanning Microscopy Based on Double-Helix Point Spread Function Engineering

Suxia Ren1, Chenshuang Zhang1, Huiqun Cao2, Danying Lin1, Bin Yu1、*, and Junle Qu1
Author Affiliations
  • 1College of Physics and Optoelectronic Engineering, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, Shenzhen University, Shenzhen 518060, Guangdong , China
  • 2College of Chemistry and Environmental Engineering, Shenzhen University, Shenzhen 518060, Guangdong , China
  • show less

    We present a method and a system of multifocal two-photon laser scanning microscopy (MTPLSM) based on double-helix point spread function (DH-PSF) engineering (DH-MTPLSM) to improve the imaging speed and resolution. In the excitation light path, a high-speed phase-only spatial light modulator (SLM) is employed to generate three-dimensional (3D) multifocal arrays and implement high-precision parallel digital addressing scanning of the sample surfaces. In the detection light path, a double-helix phase plate is inserted to modulate the system detection PSF to DH-PSF, which provides axial information of the sample. The aim is to reduce the number of layers scanned axially and ultimately enhance the speed of 3D imaging. Moreover, with a DH-PSF-based digital refocusing algorithm, we reconstruct the wide-field images of the sample at different depths and obtain the 3D optical sections of the sample by single 2D scanning. On this basis, we build a DH-MTPLSM system, carry out two-photon imaging experiments on mouse kidney sections with this system, and verify the ability of the proposed method in fast 3D high-resolution imaging, which is of great significance for the development of MTPLSM.

    Tools

    Get Citation

    Copy Citation Text

    Suxia Ren, Chenshuang Zhang, Huiqun Cao, Danying Lin, Bin Yu, Junle Qu. Three-Dimensional Multifocal Two-Photon Laser Scanning Microscopy Based on Double-Helix Point Spread Function Engineering[J]. Acta Optica Sinica, 2022, 42(14): 1411001

    Download Citation

    EndNote(RIS)BibTexPlain Text
    Save article for my favorites
    Paper Information

    Category: Imaging Systems

    Received: Nov. 26, 2021

    Accepted: Jan. 29, 2022

    Published Online: Jul. 15, 2022

    The Author Email: Yu Bin (yubin@szu.edu.cn)

    DOI:10.3788/AOS202242.1411001

    Topics