瑞典皇家理工学院Ilaria Testa小组开发出多片可逆可饱和光学荧光转换（RESOLFT）显微镜。这一研究成果于2024年2月23日在线发表在国际学术期刊《自然—方法学》上。

研究人员利用可逆转换荧光蛋白（RSFP）和周期性光模式进行光开关，展示了一种用于亚细胞结构快速容积成像的超分辨率成像方法——多片RESOLFT。通过提高记录速度（1-2Hz）并行创建宽度远低于衍射极限的多个发射片，从而提供超切片能力（<100nm）。

该技术与各种RSFP兼容，对开关周期次数的要求极低，可用于研究大量细胞结构。研究人员以三维方式跟踪细胞分裂、肌动蛋白运动和类病毒颗粒动态等细胞过程。

研究人员表示，光片荧光显微镜是对多细胞系统进行四维生物成像的宝贵工具，因为它能快速进行体积成像，而且照明剂量极小。然而，由于光片的宽度（>1μm），要检索精细的亚细胞信息，尤其是活细胞中的亚细胞信息，具有很大的挑战性。

附：英文原文

Title: Super-sectioning with multi-sheet reversible saturable optical fluorescence transitions (RESOLFT) microscopy

Author: Bodn, Andreas, Ollech, Dirk, York, Andrew G., Millett-Sikking, Alfred, Testa, Ilaria

Issue&Volume: 2024-02-23

Abstract: Light-sheet fluorescence microscopy is an invaluable tool for four-dimensional biological imaging of multicellular systems due to the rapid volumetric imaging and minimal illumination dosage. However, it is challenging to retrieve fine subcellular information, especially in living cells, due to the width of the sheet of light (>1μm). Here, using reversibly switchable fluorescent proteins (RSFPs) and a periodic light pattern for photoswitching, we demonstrate a super-resolution imaging method for rapid volumetric imaging of subcellular structures called multi-sheet RESOLFT. Multiple emission-sheets with a width that is far below the diffraction limit are created in parallel increasing recording speed (1–2Hz) to provide super-sectioning ability (<100nm). Our technology is compatible with various RSFPs due to its minimal requirement in the number of switching cycles and can be used to study a plethora of cellular structures. We track cellular processes such as cell division, actin motion and the dynamics of virus-like particles in three dimensions.

DOI: 10.1038/s41592-024-02196-8

Source: https://www.nature.com/articles/s41592-024-02196-8