Real-time multi-angle projection imaging of biological dynamics

Multiangle projection imaging accelerates volumetric imaging by up to two orders of magnitude and is readily implemented on diverse microscopes, including spinning disk confocal and light-sheet microscopes. We introduce a cost-effective and easily implementable scan unit that converts any camera-based microscope with optical sectioning capability into a multi-angle projection imaging system. Projection imaging reduces data overhead and accelerates imaging by a factor of >100, while also allowing users to readily view biological phenomena of interest from multiple perspectives on the fly. By rapidly interrogating the sample from just two perspectives, our method also enables real-time stereoscopic imaging and three-dimensional particle localization. We demonstrate projection imaging with spinning disk confocal, lattice light-sheet, multidirectional illumination light-sheet and oblique plane microscopes on specimens that range from organelles in single cells to the vasculature of a zebrafish embryo. Furthermore, we leverage our projection method to rapidly image cancer cell morphodynamics and calcium signaling in cultured neurons at rates up to 119 Hz as well as to simultaneously image orthogonal views of a beating embryonic zebrafish heart.

Automated summary

Multiangle projection imaging accelerates volumetric imaging by converting any camera-based microscope with optical sectioning capability into a system that can capture images from multiple perspectives. This method reduces data overhead, speeds up imaging, and allows for real-time stereoscopic imaging and three-dimensional particle localization. The technology has been successfully demonstrated on various microscopes and specimens, showing potential applications in cancer cell morphodynamics, calcium signaling, and simultaneous imaging of multiple perspectives of a beating heart.