Multimodal multiphoton microscopy

François Légaré; Christian P. Pfeffer M.D.; Feruz Ganikhanov

doi:10.1117/12.809096

13 February 2009 Multimodal multiphoton microscopy

François Légaré, Christian P. Pfeffer M.D., Feruz Ganikhanov

Proceedings Volume 7183, Multiphoton Microscopy in the Biomedical Sciences IX; 71831U (2009) https://doi.org/10.1117/12.809096
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Multiphoton microscopy is a powerful technique for high spatial resolution thick tissue imaging. In its simple version, it uses a high repetition rate femtosecond oscillator laser source focussed and scanned across biological sample that contains fluorophores. However, not every biological structure is inherently fluorescent or can be stained without causing biochemical changes. To circumvent these limitations, other non-invasive nonlinear optical imaging approaches are currently being developed and investigated with regard to different applications. These techniques are: (1) second harmonic generation (SHG), (2) third harmonic generation (THG), and (3) coherent anti-Stokes Raman scattering (CARS) microscopy. The main advantage of the above mentioned techniques is that they derive their imaging contrast from optical nonlinearities that do not involve fluorescence process. As a particular application example we investigated collagen arrays. We show that combining SHG-THG-CARS onto a single imaging platform provides complementary information about the sub-micron architecture of the tissue. SHG microscopy reveals the fibrillar architecture of collagen arrays and confirm a rather high degree of heterogeneity of χ⁽²⁾ within the focal volume, THG highlights the boundaries between the collagen sheets, and CH₂ spectroscopic contrast with CARS.

Citation Download Citation

François Légaré, Christian P. Pfeffer M.D., and Feruz Ganikhanov "Multimodal multiphoton microscopy", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 71831U (13 February 2009); https://doi.org/10.1117/12.809096

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