利用具有二进制数字微镜器件全息术的时聚焦多光子激发显微镜术实现快速且改进的生物成像。

Fast and improved bioimaging via temporal focusing multiphoton excitation microscopy with binary digital-micromirror-device holography.

机构信息

National Cheng Kung University, Department of Engineering Science, Tainan, Taiwan.

National Cheng Kung University, Advanced Optoelectronic Technology Center, Tainan, Taiwan.

出版信息

J Biomed Opt. 2018 Nov;23(11):1-8. doi: 10.1117/1.JBO.23.11.116502.

DOI:10.1117/1.JBO.23.11.116502

PMID:30444085

Abstract

Conventional temporal focusing-based multiphoton excitation microscopy (TFMPEM) can offer widefield optical sectioning with an axial excitation confinement of a few microns. To improve the axial confinement of TFMPEM, a binary computer-generated Fourier hologram (CGFH) via a digital-micromirror-device (DMD) was implemented to intrinsically improve the axial confinement by filling the back-focal aperture of the objective lens. Experimental results show that the excitation focal volume can be condensed and the axial confinement improved about 24% according to the DMD holography. In addition, pseudouniform MPE can be achieved using two complementary CGFHs with rapid pulse-width modulation switching via the DMD. Furthermore, bioimaging of CV-1 in origin with SV40 genes-7 cells demonstrates that the TFMPEM with binary DMD holography can improve image quality by enhancing axial excitation confinement and rejecting out-of-focus excitation.

摘要

基于传统的时间聚焦多光子激发显微镜（TFMPEM）可以提供具有几微米轴向激发限制的宽场光学切片。为了提高 TFMPEM 的轴向限制，通过数字微镜器件（DMD）实现了二元计算机生成傅里叶全息图（CGFH），通过填充物镜的后焦孔径从本质上提高了轴向限制。实验结果表明，根据 DMD 全息术，激发焦体可以被浓缩，轴向限制可以提高约 24%。此外，通过 DMD 进行快速脉宽调制切换，可以使用两个互补的 CGFH 来实现伪均匀 MPE。此外，用 SV40 基因-7 细胞转染的 CV-1 进行的生物成像表明，具有二元 DMD 全息术的 TFMPEM 可以通过增强轴向激发限制和消除离焦激发来提高图像质量。