Hyperspectral microscopic imaging by multiplex coherent anti-Stokes Raman scattering (CARS)

Alexander Khmaladze; Joshua Jasensky; Chi Zhang; Xiaofeng Han; Jun Ding; Emily Seeley; Xinran Liu; Gary D Smith; Zhan Chen

doi:10.1117/12.894258

6 September 2011 Hyperspectral microscopic imaging by multiplex coherent anti-Stokes Raman scattering (CARS)

Alexander Khmaladze, Joshua Jasensky, Chi Zhang, Xiaofeng Han, Jun Ding, Emily Seeley, Xinran Liu, Gary D Smith, Zhan Chen

Author Affiliations +

Proceedings Volume 8158, Imaging Spectrometry XVI; 815805 (2011) https://doi.org/10.1117/12.894258
Event: SPIE Optical Engineering + Applications, 2011, San Diego, California, United States

Abstract

Coherent anti-Stokes Raman scattering (CARS) microscopy is a powerful technique to image the chemical composition of complex samples in biophysics, biology and materials science. CARS is a four-wave mixing process. The application of a spectrally narrow pump beam and a spectrally wide Stokes beam excites multiple Raman transitions, which are probed by a probe beam. This generates a coherent directional CARS signal with several orders of magnitude higher intensity relative to spontaneous Raman scattering. Recent advances in the development of ultrafast lasers, as well as photonic crystal fibers (PCF), enable multiplex CARS. In this study, we employed two scanning imaging methods. In one, the detection is performed by a photo-multiplier tube (PMT) attached to the spectrometer. The acquisition of a series of images, while tuning the wavelengths between images, allows for subsequent reconstruction of spectra at each image point. The second method detects CARS spectrum in each point by a cooled coupled charged detector (CCD) camera. Coupled with point-by-point scanning, it allows for a hyperspectral microscopic imaging. We applied this CARS imaging system to study biological samples such as oocytes.

Citation Download Citation

Alexander Khmaladze, Joshua Jasensky, Chi Zhang, Xiaofeng Han, Jun Ding, Emily Seeley, Xinran Liu, Gary D Smith, and Zhan Chen "Hyperspectral microscopic imaging by multiplex coherent anti-Stokes Raman scattering (CARS)", Proc. SPIE 8158, Imaging Spectrometry XVI, 815805 (6 September 2011); https://doi.org/10.1117/12.894258

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