15 March 2018 Highly integrated label-free multiphoton nonlinear optical microspectroscopy imaging system for biomolecular imaging (Conference Presentation)
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Abstract
A biological sample consists of a variety of complex biomolecules, and fluorescence microscopy enables visualization of specific molecules at the sub-cellular level. However, these fluorescence techniques require certain fluorescence dyes to label the sample, and the fluorophores raise serious problems such as photo toxicity and photobleaching which could affect biological functionality in living systems. Advanced label-free optical imaging techniques based on nonlinear optical phenomena overcome these limitations of fluorescence microscopy. We have developed a novel label-free multimodal multiphoton nonlinear optical imaging system based on a near-IR femtosecond laser with photonic crystal fiber and pulse shaper. This highly integrated system offers numerous label-free techniques including third harmonic generation, three-photon excited fluorescence, second harmonic generation, two-photon excited fluorescence, fluorescence lifetime imaging, and broadband coherent anti-Stokes Raman scattering microspectroscopy in one platform. All of the nonlinear signals are spectrally separated by dichroic filters and simultaneously measured by photomultiplier tubes. Moreover, this system includes phase-variance optical coherence tomography as well to enable vascular imaging. We have applied our system to investigate processes in numerous biological samples. Our imaging technique is highly integrated and time efficient to generate big data, offering an array of biomolecular information at one time without staining, three-dimensional sub-micron resolution with deeper penetration, and less photodamage. The big data output from this system is analyzed by multivariate analysis such as principal component analysis and hierarchical cluster analysis. Therefore, this novel technology and methodology will have a great impact on fast in vivo label-free biomedical imaging as a big data generator.
Conference Presentation
© (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Jang Hyuk Lee, Jang Hyuk Lee, Jose Rico-Jimenez, Jose Rico-Jimenez, Aneesh Alex, Aneesh Alex, Eric J. Chaney, Eric J. Chaney, Ronit Barkalifa, Ronit Barkalifa, Haohua Tu, Haohua Tu, Marina Marjanovic, Marina Marjanovic, Zane A. Arp, Zane A. Arp, Stephen A. Boppart, Stephen A. Boppart, "Highly integrated label-free multiphoton nonlinear optical microspectroscopy imaging system for biomolecular imaging (Conference Presentation)", Proc. SPIE 10505, High-Speed Biomedical Imaging and Spectroscopy III: Toward Big Data Instrumentation and Management, 105050V (15 March 2018); doi: 10.1117/12.2287194; https://doi.org/10.1117/12.2287194
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