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7 March 2018 Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation
Andrea Filippi, Eleonora Dal Sasso, Laura Iop, Andrea Armani, Michele Gintoli, Marco Sandri, Gino Gerosa, Filippo Romanato, Giulia Borile
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Journal of Biomedical Optics, Vol. 23, Issue 9, 091403 (March 2018). https://doi.org/10.1117/1.JBO.23.9.091403
Abstract
Label-free microscopy is a very powerful technique that can be applied to study samples with no need for exogenous fluorescent probes, keeping the main benefits of multiphoton microscopy, such as longer penetration depths and intrinsic optical sectioning while enabling serial multitechniques examinations on the same specimen. Among the many label-free microscopy methods, harmonic generation (HG) is one of the most intriguing methods due to its generally low photo-toxicity and relative ease of implementation. Today, HG and common two-photon microscopy (TPM) are well-established techniques, and are routinely used in several research fields. However, they require a significant amount of fine-tuning to be fully exploited, making them quite difficult to perform in parallel. Here, we present our designed multimodal microscope, capable of performing simultaneously TPM and HG without any kind of compromise thanks to two, separate, individually optimized laser sources with axial chromatic aberration compensation. We also apply our setup to the examination of a plethora of ex vivo samples to prove its capabilities and the significant advantages of a multimodal approach.
© 2018 Society of Photo-Optical Instrumentation Engineers (SPIE) 1083-3668/2018/$25.00 © 2018 SPIE
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Andrea Filippi, Eleonora Dal Sasso, Laura Iop, Andrea Armani, Michele Gintoli, Marco Sandri, Gino Gerosa, Filippo Romanato, and Giulia Borile "Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation," Journal of Biomedical Optics 23(9), 091403 (7 March 2018). https://doi.org/10.1117/1.JBO.23.9.091403
Received: 25 November 2017; Accepted: 12 February 2018; Published: 7 March 2018
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KEYWORDS
Second-harmonic generation
Signal detection
Collagen
Chromatic aberrations
Polarization
Microscopes
Microscopy
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