Your browser doesn't support javascript.
loading
Super-resolution SRS microscopy with A-PoD.
Jang, Hongje; Li, Yajuan; Fung, Anthony A; Bagheri, Pegah; Hoang, Khang; Skowronska-Krawczyk, Dorota; Chen, Xiaoping; Wu, Jane Y; Bintu, Bogdan; Shi, Lingyan.
Affiliation
  • Jang H; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Li Y; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Fung AA; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Bagheri P; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Hoang K; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Skowronska-Krawczyk D; School of Medicine, University of California, Irvine, CA, USA.
  • Chen X; The Ken and Ruth Davee Department of Neurology, Northwestern University, Chicago, IL, USA.
  • Wu JY; The Ken and Ruth Davee Department of Neurology, Northwestern University, Chicago, IL, USA.
  • Bintu B; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
  • Shi L; Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA. lingyanshi@ucsd.edu.
Nat Methods ; 20(3): 448-458, 2023 03.
Article in En | MEDLINE | ID: mdl-36797410
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.
Subject(s)
Fulltext
Add to My VHL
Print
XML
PubMed Links
Search on Google

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Spectrum Analysis, Raman / Microscopy Language: En Journal: Nat Methods Journal subject: TECNICAS E PROCEDIMENTOS DE LABORATORIO Year: 2023 Document type: Article Affiliation country: United States Country of publication: United States
Fulltext
Add to My VHL
Print
XML
PubMed Links
Search on Google

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Spectrum Analysis, Raman / Microscopy Language: En Journal: Nat Methods Journal subject: TECNICAS E PROCEDIMENTOS DE LABORATORIO Year: 2023 Document type: Article Affiliation country: United States Country of publication: United States