Expansion and Light-Sheet Microscopy for Nanoscale 3D Imaging

Expansion Microscopy (ExM) and Light-Sheet Fluorescence Microscopy (LSFM) are forefront imaging techniques that enable high-resolution visualization of biological specimens. ExM enhances nanoscale investigation using conventional fluorescence microscopes, while LSFM offers rapid, minimally invasive imaging over large volumes. This review explores the joint advancements of ExM and LSFM, focusing on the excellent performance of the integrated modality obtained from the combination of the two, which is refer to as ExLSFM. In doing so, the chemical processes required for ExM, the tailored optical setup of LSFM for examining expanded samples, and the adjustments in sample preparation for accurate data collection are emphasized. It is delve into various specimen types studied using this integrated method and assess its potential for future applications. The goal of this literature review is to enrich the comprehension of ExM and LSFM, encouraging their wider use and ongoing development, looking forward to the upcoming challenges, and anticipating innovations in these imaging techniques. Expansion Microscopy (ExM) and Light-Sheet Fluorescence Microscopy (LSFM) converge in ExLSFM, blending ExM's nanoscale precision with LSFM's rapid imaging of large volumes. The review underscores ExM's chemical processes, LSFM's optical setup for expanded samples, and sample preparation. Examining diverse specimen types, it reports ExLSFM's versatile applications. The goal is to enhance comprehension, foster wider adoption, and foresee innovations and challenges in ExM and LSFM. image