Gialdini Irene, Hendrix Jelle, Lamb Don C
Department Chemie and Center for NanoScience, Ludwig-Maximilians-Universität München, Munich, Germany.
Dynamic Bioimaging Lab, Advanced Optical Microscopy Centre and Biomedical Research Institute, Hasselt University, Agoralaan C (BIOMED), Hasselt, Belgium.
Biochim Biophys Acta Gen Subj. 2025 Jul;1869(8):130818. doi: 10.1016/j.bbagen.2025.130818. Epub 2025 May 10.
Raster Image Correlation Spectroscopy (RICS) is a confocal image analysis method that can measure the diffusion and interactions of fluorescently labeled molecules in real time in solution and in living cells. RICS is easy to implement on commercial confocal microscopes and allows detailed investigations of complex biological systems and pathways. The method is especially robust for measurements in living cells using commonly used labels such as fluorescent proteins. Moreover, since its invention in 2005, the robustness and applicability of RICS has been significantly increased to allow, e.g., straightforward kinetic analyses, advanced image segmentation, parameter mapping, and multi-species analysis. In this review, we describe the methodological principles of RICS in a manner that is accessible to a broad readership, position RICS in relation to other fluorescence fluctuation techniques, highlight recent methodological advances and present exemplary applications of the method. With this review, we hope to facilitate the implementation of this powerful method into the everyday repertoire of confocal imaging approaches.
光栅图像相关光谱法（RICS）是一种共聚焦图像分析方法，可实时测量溶液和活细胞中荧光标记分子的扩散及相互作用。RICS易于在商用共聚焦显微镜上实施，能够对复杂的生物系统和信号通路进行详细研究。该方法对于使用荧光蛋白等常用标记物在活细胞中进行测量特别可靠。此外，自2005年发明以来，RICS的可靠性和适用性已显著提高，例如可进行直接的动力学分析、先进的图像分割、参数映射和多物种分析。在本综述中，我们以广大读者易于理解的方式描述RICS的方法原理，将RICS与其他荧光波动技术进行定位比较，突出近期的方法进展，并展示该方法的典型应用。通过本综述，我们希望促进将这种强大的方法应用于共聚焦成像方法的日常操作中。
