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Updated: Sep 13, 2025

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
Published on: April 7, 2014
Snapshot Quantitative Phase Imaging with Acousto-Optic Chromatic Aberration Control
Christos Alexandropoulos1, Laura Rodríguez-Suñé1, Martí Duocastella1,2
1Department of Applied Physics, Universitat de Barcelona, C/Martí i Franquès 1, 08028 Barcelona, Spain.
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The transport of intensity equation enables quantitative phase imaging from only two axially displaced intensity images, facilitating the characterization of low-contrast samples like cells and microorganisms. However, the rapid selection of the correct defocused planes, crucial for real-time phase imaging of dynamic events, remains challenging. Additionally, the different images are normally acquired sequentially, further limiting phase-reconstruction speed. Here, we report on a system that addresses these issues and enables user-tuned defocusing with snapshot phase retrieval. Our approach is based on combining multi-color pulsed illumination with acousto-optic defocusing for microsecond-scale chromatic aberration control. By illuminating each plane with a different color and using a color camera, the information to reconstruct a phase map can be gathered in a single acquisition. We detail the fundamentals of our method, characterize its performance, and demonstrate live phase imaging of a freely moving microorganism at speeds of 150 phase reconstructions per second, limited only by the camera's frame rate.

