Coupling CARS with multiphoton fluorescence and high harmonic generation imaging modalities using a femtosecond laser source

Hongtao Chen; Mikhail N. Slipchenko; Jiabin Zhu; Kimberly K. Buhman; Ji-Xin Cheng D.V.M.

doi:10.1117/12.808223

13 February 2009 Coupling CARS with multiphoton fluorescence and high harmonic generation imaging modalities using a femtosecond laser source

Hongtao Chen, Mikhail N. Slipchenko, Jiabin Zhu, Kimberly K. Buhman, Ji-Xin Cheng D.V.M.

Author Affiliations +

Proceedings Volume 7183, Multiphoton Microscopy in the Biomedical Sciences IX; 718315 (2009) https://doi.org/10.1117/12.808223
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Multimodal nonlinear optical imaging has opened new opportunities and becomes a powerful tool for imaging complex tissue samples with inherent 3D spatial resolution.. We present a robust and easy-to-operate approach to add the coherent anti-stokes Raman scattering (CARS) imaging modality to a widely used multiphoton microscope. The laser source composed of a Mai Tai femtosecond laser and an optical parametric oscillator (OPO) offers one-beam, two-beam and three-beam modalities. The Mai Tai output at 790 nm is split into two beams, with 80% of the power being used to pump the OPO. The idler output at 2036 nm from OPO is doubled using a periodically poled lithium niobate (PPLN) crystal. This frequency-doubled idler beam at 1018 nm is sent through a delay line and collinearly combined with the other Mai Tai beam for CARS imaging on a laser-scanning microscope. This Mai Tai beam is also used for multiphoton fluorescence and second harmonic generation (SHG) imaging. The signal output at 1290 nm from OPO is used for SHG and third-harmonic generation (THG) imaging. External detectors are installed for both forward and backward detection, whereas two internal lamda-scan detectors are employed for microspectroscopy analysis. This new system allows vibrationally resonant CARS imaging of lipid bodies, SHG imaging of collagen fibers, and multiphoton fluorescence analysis in fresh tissues. As a preliminary application, the effect of diacylglycerol acyltransferase 1 (DGAT1) deficiency on liver lipid metabolism in mice was investigated.

Citation Download Citation

Hongtao Chen, Mikhail N. Slipchenko, Jiabin Zhu, Kimberly K. Buhman, and Ji-Xin Cheng D.V.M. "Coupling CARS with multiphoton fluorescence and high harmonic generation imaging modalities using a femtosecond laser source", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 718315 (13 February 2009); https://doi.org/10.1117/12.808223

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