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Highly-Multiplexed Tissue Imaging with Raman Dyes
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JoVE Journal Bioingegneria
Highly-Multiplexed Tissue Imaging with Raman Dyes

Highly-Multiplexed Tissue Imaging with Raman Dyes

DOI:
10.3791/63547-v

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07:18 min

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April 21, 2022

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, ,
1Department of Chemistry,Columbia University, 2Kavli Institute for Brain Science,Columbia University

Capitoli

  • 00:04Introduction
  • 00:59Preparation of Raman-Dye-Conjugated Antibodies
  • 02:57Tissue Immuno-eprSRS Staining
  • 05:02Image Acquisition and Analysis
  • 05:35Results: Electronic Pre-Resonance Stimulated Raman Scattering Imaging of Raman Dyes
  • 06:49Conclusion

Summary

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Electronic pre-resonance stimulated Raman scattering (epr-SRS) imaging of rainbow-like Raman dyes is a new platform for highly multiplexed epitope-based protein imaging. Here, we present a practical guide including antibody preparation, tissue sample staining, SRS microscope assembly, and epr-SRS tissue imaging.

Tags

Keywords: Highly-multiplexedVibrational ImagingProtein MarkersSubcellular ResolutionCycle-free MultiplexityCell CharacterizationTumor MicroenvironmentsBrain CircuitSodium BicarbonatePBS BufferN-hydroxysuccinimide EsterMARS ProbeAntibody ConjugationSize Exclusion ColumnPurification

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