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High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
Published on: July 25, 2014
Time-of-flight fluorescence depth mapping using a spatiotemporal deep learning model
Shiru Wang1, Arthur Pétusseau1, Claudio Bruschini2
1Dartmouth College, Thayer School of Engineering, Hanover, New Hampshire United States.
Journal of Biomedical Optics
|June 22, 2026
Summary
This study introduces a deep learning method using SPAD sensor images to map subsurface fluorescence depth, improving surgical precision. The system accurately reconstructs fluorescence topography, overcoming optical artifacts in real-time for enhanced surgical guidance.
Area of Science:
- Medical Imaging
- Biophotonics
- Artificial Intelligence
Background:
- Fluorescence-guided surgery (FGS) uses molecular contrast agents for real-time visualization but is limited by optical artifacts like diffusion and absorption.
- These artifacts hinder accurate quantitative assessment of fluorophore distribution, potentially compromising surgical precision.
- Accurate sub-surface fluorescence imaging requires resolving topological depth maps at the millimeter scale.
Purpose of the Study:
- To develop a spatiotemporal deep learning architecture for rapid recovery of fluorophore depth topology using picosecond single-photon avalanche diode (SPAD) sensor images.
- To enable precise surgical decision-making by overcoming limitations in current FGS techniques.
- To achieve millimeter spatial resolution in wide-field, epi-illumination geometry for sub-surface fluorescence imaging.
Main Methods:
- A ConvLSTM-UNet deep learning network was developed for analyzing picosecond time-resolved images.
- The network was trained on simulated spatiotemporal maps using optical Monte Carlo methods and the instrument response function (IRF).
- The system utilized a SwissSPAD2 sensor synchronized with a picosecond laser diode, processing data from 10 temporal gates.
Main Results:
- The deep learning network successfully reconstructed fluorescence topography up to 15 mm depth.
- Reconstruction accuracy achieved a mean absolute error of less than 0.6 mm and mean depth variances below 0.5 mm.
- The system demonstrated a rapid inference time of approximately 30 ms.
Conclusions:
- Integrating temporal and spatial deep learning networks enables accurate depth mapping from time-resolved fluorescence data.
- The use of real IRF confirmed the applicability of SPAD sensors for sub-surface fluorescence mapping.
- This approach enhances the capabilities of fluorescence-guided surgery by providing quantitative depth information.
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