Super-resolution SRS microscopy with A-PoD

Hongje Jang, Yajuan Li, Anthony A. Fung, Pegah Bagheri, Khang Hoang, Dorota Skowronska-Krawczyk, Xiaoping Chen, Jane Y. Wu, Bogdan Bintu, Lingyan Shi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Stimulated Raman scattering (SRS) offers the ability to image metabolic dynamics with high signal-to-noise ratio. However, its spatial resolution is limited by the numerical aperture of the imaging objective and the scattering cross-section of molecules. To achieve super-resolved SRS imaging, we developed a deconvolution algorithm, adaptive moment estimation (Adam) optimization-based pointillism deconvolution (A-PoD) and demonstrated a spatial resolution of lower than 59 nm on the membrane of a single lipid droplet (LD). We applied A-PoD to spatially correlated multiphoton fluorescence imaging and deuterium oxide (D2O)-probed SRS (DO-SRS) imaging from diverse samples to compare nanoscopic distributions of proteins and lipids in cells and subcellular organelles. We successfully differentiated newly synthesized lipids in LDs using A-PoD-coupled DO-SRS. The A-PoD-enhanced DO-SRS imaging method was also applied to reveal metabolic changes in brain samples from Drosophila on different diets. This new approach allows us to quantitatively measure the nanoscopic colocalization of biomolecules and metabolic dynamics in organelles.

Original languageEnglish (US)
Pages (from-to)448-458
Number of pages11
JournalNature Methods
Issue number3
StatePublished - Mar 2023

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry
  • Biotechnology
  • Cell Biology


Dive into the research topics of 'Super-resolution SRS microscopy with A-PoD'. Together they form a unique fingerprint.

Cite this