Super-resolution SRS microscopy with A-PoD

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.