Advances in Rolling Shutter Line-Scan Microscopy

Recent advancements in 3D fluorescence microscopy have significantly enhanced the ability to study biological processes in vivo with high resolution. However, the complexity and cost associated with state-of-the-art systems like lattice lightsheet microscopy and oblique plane microscopy often limit their accessibility to specialized laboratories. This Review explores an emerging class of microscopy techniques that utilize line-scan illumination combined with rolling shutter detection, offering a simpler and more accessible alternative that still maintains high imaging performance. These methods extend the principles of traditional confocal microscopy by illuminating the sample along a line and using a camera’s rolling shutter as a virtual slit to enhance optical sectioning and resolution. Various implementations are discussed to highlight their unique benefits and applications. In addition, this Review provides a comprehensive comparison between implementations and showcases latest systems that gain unique capabilities enabled via clever use of rolling shutter line-scanning. By making high-resolution imaging more accessible, rolling shutter line-scan microscopy is prone to empower a wide range of scientists to explore intricate cellular dynamics and structures, paving the way for new discoveries in the life sciences.