This paper revives the remote focusing technique by presenting a new device, allowing to acquire fast, high resolution 3D images in fluorescence microscopy, in order to reveal 3D dynamics of living samples.
By imaging many Argo-SIM fluorescent test patterns, both 2D and 3D (the last is a unique capability of Argolight’s technology), the authors, from Calico Life Sciences LLC (a subsidiary of Google), demonstrate the performances of their remote refocus device: Camera-limited speed and diffraction-limited resolution over a user-specified volume up 200×200×60 µm3.
A. Millett-Sikking, N. H. Thayer, A. Bohnert and A. G. York, “Remote refocus enables class-leading spatiotemporal resolution in 4D optical microscopy,” hosted on GitHub Pages, DOI:10.5281/zenodo.1146083, January 2018.
Volumetric imaging capability of remote refocus illustrated via 3D structures on the Argolight SIM slide. The pattern consists of a 3D array of 9x9x9 rings separated by 5 μm in X, Y, and Z. We acquired the data by focusing the microscope Z-drive onto the top plane, and then used the RR piezo to take the Z-stack. We processed the stack with ImageJ to give 3D projections from various angles. Scale bar is 20 μm.