Gimeno-Gomez Alejandro, Dajkhosh Seyedeh Parisa, Son Van Cong Tuan, Barreiro Juan Carlos, Preza Chrysanthe, Saavedra Genaro
Opt Express. 2023 Oct 23;31(22):36568-36589. doi: 10.1364/OE.496518.
We have designed and implemented an approach for three-dimensional (3D) structured illumination (SI) microscopy (SIM) based on a quasi-monochromatic extended source illuminating a Wollaston prism to improve robustness, light efficiency and flexibility over our previous design. We show through analytical and experimental verification of the presented theoretical framework for our proposed tunable structured illumination microscopy (TSIM) system, that a simple and accurate determination of the axial modulation of the SI pattern is achieved, enabling a realistic characterization of the system's effective optical transfer function (OTF). System performance as a function of the extended source size is investigated with simulations. Results from a comparative performance analysis of the proposed TSIM system and traditional SIM systems show some advantages over the traditional two-wave and three-wave interference SIM systems. We show that by controlling the source size and thereby the axial modulation of the 3D SI pattern, the TSIM scheme offers increased OTF compact support and improved optical sectioning capability, quantified by the integrated intensity, under certain conditions, which may be desirable when imaging optically thick samples. The additional tunability of the 3D SI pattern, provides a unique opportunity for OTF engineering in our TSIM system.
我们设计并实现了一种基于准单色扩展光源照射沃拉斯顿棱镜的三维（3D）结构照明（SI）显微镜（SIM）方法，以提高其相对于我们之前设计的鲁棒性、光效率和灵活性。通过对我们提出的可调谐结构照明显微镜（TSIM）系统的理论框架进行分析和实验验证，我们表明可以简单而准确地确定SI图案的轴向调制，从而能够真实地表征系统的有效光学传递函数（OTF）。通过模拟研究了系统性能与扩展光源大小的关系。对所提出的TSIM系统与传统SIM系统的比较性能分析结果表明，该系统相对于传统的两波和三波干涉SIM系统具有一些优势。我们表明，通过控制光源大小从而控制3D SI图案的轴向调制，TSIM方案在某些条件下提供了更大的OTF紧凑支持并提高了光学切片能力，这可以通过积分强度来量化，在对光学厚样品成像时可能是理想的。3D SI图案的额外可调性为我们的TSIM系统中的OTF工程提供了独特的机会。
