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Updated: Apr 28, 2026

Registration of Calcium Transients in Mouse Neuromuscular Junction with High Temporal Resolution using Confocal Microscopy
Published on: December 1, 2021
Scanless temporal focusing enables high-speed three-dimensional quantitative phase microscopy
Peter So1,2, Yuechuan Lin2,3, Xiang Zhang2,3
1Laser Biomedical Research Center, G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
None:
Quantitative phase microscopy (QPM) enables label-free imaging of structure and dynamics in biological and physical systems, yet achieving high-speed three-dimensional (3D) QPM with strong optical sectioning remains a central challenge. Here, we introduce a single-shot reflection-mode temporal focusing QPM (TF-QPM) that provides sub-micron optical sectioning without needs of any mechanical scanning or multiplexed acquisitions. By extending temporal focusing beyond its conventional use in multiphoton fluorescence microscopy, TF-QPM enables diffraction-limited label-free phase-sensitive volumetric imaging with 402 nm lateral and 920 nm axial resolution, markedly reduced speckle noise, and depth-resolved imaging at 3,709 Hz frame rate -an order of magnitude faster than most existing techniques and currently only limited by the camera speed. The resulting spatiotemporal phase sensitivity enables precise 3D tracking of particle motion and quantitative characterization of fast dynamics in complex and anisotropic media. For tissue imaging applications, TF-QPM achieves histology-level resolution in intact samples and supports pixel-level virtual staining, providing a rapid, label-free alternative to conventional sectioning-based workflows. Together, these results establish TF-QPM as a scanless, high-speed platform for rapid, label-free volumetric imaging across both basic research and translational applications.

