Home › Analytical Chem › Differential Imaging of Biological Structures with Doubly-resonant Coherent Anti-stokes Raman Scattering (CARS)
Analytical Chem JoVE (Open Access) Citable · DOI

Differential Imaging of Biological Structures with Doubly-resonant Coherent Anti-stokes Raman Scattering (CARS)

DOI: 10.3791/2085-v
What you'll learn
  • ✓ Understand the principles of coherent anti-Stokes Raman scattering (CARS) and doubly-resonant CARS
  • ✓ Operate a multi-laser DR-CARS microscopy system for biological imaging
  • ✓ Prepare biological samples and acquire high-sensitivity Raman images of weak scatterers
Protocol

A combination of three single wavelength short-pulsed lasers is used to generate coherent anti-Stokes Raman scattering (CARS) and doubly-resonant CARS (DR-CARS). The difference between these signals provides enhanced sensitivity for otherwise difficult to detect coherent Raman signals, enabling imaging of weak Raman scatterers.

Difficulty
advanced
Total time
~2–4 hours per sample (including system alignment and image acquisition)

Steps

1
Understand CARS and DR-CARS signal generation principles

Review the physical basis for coherent anti-Stokes Raman scattering and how doubly-resonant CARS enhances detection sensitivity for weak Raman scatterers.

▶ 00:46
2
Configure the DR-CARS microscopy system

Set up the three single-wavelength short-pulsed laser sources and align them for coherent signal generation and biological structure imaging.

▶ 05:21
3
Prepare biological samples for DR-CARS imaging

Prepare samples according to protocol requirements, ensuring proper mounting and positioning for microscopic analysis.

▶ 07:02
4
Acquire and analyze DR-CARS images

Perform imaging with the DR-CARS system and interpret differential signal data to visualize biological structures with enhanced contrast.

▶ 11:16
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