Zapata-Valencia Samuel I, Tobón-Maya Heberley, Ordoñez Luis, Farina Andrea, Lancis Jesús, Tajahuerce Enrique
Institute of New Imaging Technologies (INIT), Universitat Jaume I, Castelló de la Plana, 12071, Spain.
Dipartimento di Fisica, Politecnico di Milano, Piazza L. da Vinci 32, Milan, 20133, Italy.
Sci Rep. 2025 May 22;15(1):17802. doi: 10.1038/s41598-025-03125-1.
Single-pixel microscopy (SPM) is an emerging imaging technique in which a sample is illuminated with a series of micro-structured light patterns, typically generated by a digital micromirror device (DMD). After interaction with the sample, light is collected by a bucket detector, and the image is reconstructed through computational algorithms, such as basis transformations or compressive sensing. DMD achievable framerates and wide spectral range has allowed SPM to develop a wide range of applications, including polarization state analysis, phase imaging, and fluorescence lifetime measurements. To achieve optimal performance in these applications, a precise system configuration is required ensuring the effective projection of the structured light patterns. Nevertheless, the incorporation of a DMD introduces additional complexity, particularly in alignment, which can significantly affect system performance if not properly addressed. This work presents a comprehensive framework for the design, assembly, and alignment of a modular, open-source SPM system. The proposed procedures minimize aberrations introduced during construction and ensures the accurate projection of structured light patterns onto the sample. The modular design facilitates integration across multiple illumination sources and enables simultaneous brightfield transmission and reflection imaging. The proposed system achieves resolution near the diffraction limit, surpassing previous SPM configurations without requiring numerical or optical enhancement techniques. Performance validation through imaging experiments on both biological and non-biological samples demonstrates the system's robustness and versatility. By providing detailed design and assembly instructions, this work contributes to the openness and reproducibility of SPM and serves as a valuable resource for researchers aiming to build high-performance, customizable single-pixel imaging systems.
单像素显微镜（SPM）是一种新兴的成像技术，在该技术中，样品由一系列微结构光图案照明，这些图案通常由数字微镜器件（DMD）生成。与样品相互作用后，光由桶形探测器收集，图像通过诸如基变换或压缩感知等计算算法重建。DMD可实现的帧率和宽光谱范围使SPM能够开发出广泛的应用，包括偏振态分析、相位成像和荧光寿命测量。为了在这些应用中实现最佳性能，需要精确的系统配置以确保结构光图案的有效投影。然而，DMD的加入带来了额外的复杂性，特别是在对准方面，如果处理不当，可能会显著影响系统性能。这项工作提出了一个用于模块化、开源SPM系统的设计、组装和对准的综合框架。所提出的程序可将构建过程中引入的像差降至最低，并确保结构光图案准确投影到样品上。模块化设计便于跨多个照明源进行集成，并能够同时进行明场透射和反射成像。所提出的系统实现了接近衍射极限的分辨率，超越了以前的SPM配置，而无需数值或光学增强技术。通过对生物和非生物样品进行成像实验进行性能验证，证明了该系统的稳健性和通用性。通过提供详细的设计和组装说明，这项工作有助于提高SPM的开放性和可重复性，并为旨在构建高性能、可定制单像素成像系统的研究人员提供了宝贵的资源。
