National Cheng Kung University, Department of Engineering Science, Tainan, Taiwan.
National Cheng Kung University, Advanced Optoelectronic Technology Center, Tainan, Taiwan.
J Biomed Opt. 2018 Nov;23(11):1-8. doi: 10.1117/1.JBO.23.11.116502.
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.
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。
