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Updated: Nov 15, 2025

Three-dimensional Optical-resolution Photoacoustic Microscopy
Published on: May 3, 2011
Live feedback and 3D photoacoustic remote sensing
Saad Abbasi1, Kevan Bell1,2, Benjamin Ecclestone1
1PhotoMedicine Labs, Department of Systems Design Engineering, University of Waterloo, Waterloo, Ontario, Canada.
Background:
As photoacoustic (PA) techniques progress towards clinical adoption, providing a high-speed live feedback becomes a high priority. To keep up with the instantaneous optical feedback of conventional light microscopes, PA imaging would need to provide a high-resolution video-rate live feed to the user. However, conventional PA microscopy typically trades resolution, sensitivity and imaging speed when optically scanning due to the difficult opto-acoustic confocal geometry. Here, we employ photoacoustic remote sensing (PARS), an all-optical technique that relies on optical confocal geometry, to provide a high-resolution live display in a reflection-mode PA architecture.
Methods:
Employing a conventional x-y galvanometer scanner and a 600 KHz pulse repetition rate laser we implement a system capable of acquiring 2.5 frames per second in 2D. To complement this fast scanning optical system, we implement a computationally inexpensive image reconstruction method that is able to render the frames with minimal overhead, providing a live display.
Results:
Employing the proposed method, we demonstrate a live feedback with frame rates as high as 2.5 Hz in 2D and also report the first results of 3D imaging with a non-contact label-free reflection-mode technique. The method is validated with phantom studies and in-vivo imaging. Employing a repetition rate of 600 KHz, a live feed of carbon fibers is realized with a C-scan rate of 2.5 Hz. The imaging resolution was measured to be 1.2 µm, the highest reported for a real-time reflection-mode architecture. The mean and peak SNR were measured to be 44 and 62 dB respectively in-vivo. 3D visualizations of carbon fiber phantoms and mouse ear microvasculature structure are also demonstrated.
Conclusions:
In summary, we present a method that has a small computational overhead for image rendering, resulting in a live display capable of real-time frame rates. We also report the first 3D imaging with a non-contact label-free reflection-mode PA technique. The all-optical confocal geometry required by PARS is significantly easier to implement and maintain than the opto-acoustic geometry of conventional PA microscopy techniques. This results in a system capable of high resolution and sensitivity, imaging at real-time rates. The authors believe this work represents a vital step towards a clinical high-resolution reflection-mode video-rate PA imaging system.
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