Significance: High-speed three-dimensional (3D) super-resolution microscopy is a unique tool to investigate various biological phenomena; yet the technology is not broadly adopted due to its high cost and complex system design. Aim: We present a compact, low-cost, and high-speed 3D structured illumination microscopy (SIM) based on a digital micromirror device and binary holography to visualize fast biological events with super-resolution. Approach: The 3D SIM uses a digital micromirror device to generate three laser foci with individually controllable positions, phases, and amplitudes via binary holography at the back aperture of objective lens to form optimal 3D structured patterns. Fifteen raw images are sequentially recorded and processed by the 3D SIM algorithm to reconstruct a super-resolved image. Results: Super-resolution 3D imaging at a speed of 26.7 frames per second is achieved with a lateral and axial resolution of 155 and 487 nm, which corresponds to a 1.65- and 1.63-times resolution enhancement, respectively, comparing with standard deconvolution microscopy. Conclusions: The 3D SIM realizes fast super-resolution imaging with optimal 3D structured illumination, which may find important applications in biophotonics.

中文翻译：

---

使用数字微镜设备和二元全息技术的快速 3D 超分辨率成像

意义：高速三维（3D）超分辨率显微镜是研究各种生物现象的独特工具；但由于成本高、系统设计复杂，该技术并未得到广泛采用。目标：我们提出了一种基于数字微镜设备和二元全息术的紧凑、低成本和高速 3D 结构照明显微镜 (SIM)，以超分辨率可视化快速生物事件。方法：3D SIM 使用数字微镜装置通过物镜后孔处的二元全息术产生位置、相位和幅度可单独控制的三个激光焦点，以形成最佳的 3D 结构图案。15 张原始图像依次记录并通过 3D SIM 算法处理以重建超分辨率图像。结果：以每秒 26.7 帧的速度实现超分辨率 3D 成像，横向和轴向分辨率分别为 155 和 487 nm，与标准去卷积显微镜相比，分辨率分别提高了 1.65 倍和 1.63 倍。结论：3D SIM 实现了具有最佳 3D 结构照明的快速超分辨率成像，这可能会在生物光子学中找到重要应用。