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

doi:10.1038/s41592-023-01779-1

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

Neurology

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

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 language	English (US)
Pages (from-to)	448-458
Number of pages	11
Journal	Nature Methods
Volume	20
Issue number	3
DOIs	https://doi.org/10.1038/s41592-023-01779-1
State	Published - Mar 2023

Funding

We thank K. Zhang, W. Min and J. Enderlein for helpful discussions and suggestions. Thanks to M. Shtrahman and S. Saidi for providing 1 µm bead samples. We acknowledge University of California, San Diego startup funds, NIH U54CA132378, NIH 5R01NS111039, NIH R21NS125395, NIH U54DK134301, NIH U54 HL165443 and a Hellman Fellow Award.

ASJC Scopus subject areas

Molecular Biology
Biochemistry
Biotechnology
Cell Biology

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title = "Super-resolution SRS microscopy with A-PoD",

abstract = "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.",

author = "Hongje Jang and Yajuan Li and Fung, {Anthony A.} and Pegah Bagheri and Khang Hoang and Dorota Skowronska-Krawczyk and Xiaoping Chen and Wu, {Jane Y.} and Bogdan Bintu and Lingyan Shi",

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AU - Skowronska-Krawczyk, Dorota

AU - Chen, Xiaoping

AU - Wu, Jane Y.

AU - Bintu, Bogdan

AU - Shi, Lingyan

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N2 - 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.

AB - 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.

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Super-resolution SRS microscopy with A-PoD

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