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Related Experiment Video

Updated: Aug 9, 2025

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

Published on: July 6, 2019

Super-resolution SRS microscopy with A-PoD.

Hongje Jang¹, Yajuan Li¹, Anthony A Fung¹

¹Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.

|February 16, 2023

Summary

This summary is machine-generated.

We developed a new super-resolution imaging technique, adaptive moment estimation (Adam) optimization-based pointillism deconvolution (A-PoD), to visualize metabolic processes at the nanoscale. This method achieves unprecedented spatial resolution for studying cellular lipid and protein dynamics.

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Last Updated: Aug 9, 2025

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

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Published on: July 6, 2019

In vivo Imaging of Biological Tissues with Combined Two-Photon Fluorescence and Stimulated Raman Scattering Microscopy

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Published on: December 20, 2021

Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy

Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy

Published on: May 16, 2022

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Stimulated Raman scattering (SRS) microscopy enables high signal-to-noise imaging of metabolic dynamics.
Current SRS spatial resolution is limited by optical factors and molecular scattering cross-sections.
Nanoscale imaging of metabolic processes is crucial for understanding cellular function.

Purpose of the Study:

To develop a super-resolution technique for SRS imaging.
To enhance the spatial resolution of SRS beyond conventional limits.
To quantitatively analyze the nanoscopic distribution and dynamics of biomolecules in cells and organelles.

Main Methods:

Development of adaptive moment estimation (Adam) optimization-based pointillism deconvolution (A-PoD) algorithm.
Application of A-PoD to spatially correlated multiphoton fluorescence and deuterium oxide (D2O)-probed SRS (DO-SRS) imaging.
Imaging of lipid droplets, cellular organelles, and Drosophila brain samples.

Main Results:

Achieved spatial resolution below 59 nm on lipid droplet membranes.
Successfully differentiated newly synthesized lipids in lipid droplets using A-PoD-coupled DO-SRS.
Revealed metabolic changes in Drosophila brain samples subjected to different diets.
Quantitatively measured nanoscopic colocalization of biomolecules and metabolic dynamics.

Conclusions:

A-PoD significantly enhances SRS spatial resolution, enabling nanoscale metabolic imaging.
The A-PoD-enhanced DO-SRS method provides new insights into lipid and protein distributions in cellular environments.
This technique is valuable for studying metabolic dynamics and molecular colocalization in various biological samples, including disease models.