Stony Brook University (SUNY), Department of Biomedical Engineering, Stony Brook, New York 11794, USA.
J Biomed Opt. 2013 Jun;18(6):066006. doi: 10.1117/1.JBO.18.6.066006.
Dual-axis confocal (DAC) microscopy has been found to exhibit superior rejection of out-of-focus and multiply scattered background light compared to conventional single-axis confocal microscopy. DAC microscopes rely on the use of separated illumination and collection beam paths that focus and intersect at a single focal volume (voxel) within tissue. While it is generally recognized that the resolution and contrast of a DAC microscope depends on both the crossing angle of the DAC beams, 2θ, and the focusing numerical aperture of the individual beams, α, a detailed study to investigate these dependencies has not been performed. Contrast and resolution are considered as two main criteria to assess the performance of a point-scanned DAC microscope (DAC-PS) and a line-scanned DAC microscope (DAC-LS) as a function of θ and α. The contrast and resolution of these designs are evaluated by Monte-Carlo scattering simulations and diffraction theory calculations, respectively. These results can be used for guiding the optimal designs of DAC-PS and DAC-LS microscopes.
双轴共焦（DAC）显微镜已被发现与传统的单轴共焦显微镜相比，具有更好的离焦和多次散射背景光的抑制能力。DAC 显微镜依赖于使用分离的照明和收集光束路径，这些光束在组织内的单个焦点体积（体素）处聚焦并相交。虽然人们普遍认为 DAC 显微镜的分辨率和对比度取决于 DAC 光束的交叉角 2θ 和单个光束的聚焦数值孔径 α，但尚未进行详细研究以调查这些依赖性。对比度和分辨率被认为是评估点扫描 DAC 显微镜（DAC-PS）和线扫描 DAC 显微镜（DAC-LS）作为 θ 和 α 的函数的性能的两个主要标准。这些设计的对比度和分辨率分别通过蒙特卡罗散射模拟和衍射理论计算进行评估。这些结果可用于指导 DAC-PS 和 DAC-LS 显微镜的最佳设计。
