High speed multiphoton imaging

Yongxiao Li; Anne Brustle; Vini Gautam; Ian Cockburn; Cathy Gillespie; Katharina Gaus; Woei Ming Lee

doi:10.1117/12.2242951

9 December 2016 High speed multiphoton imaging

Yongxiao Li, Anne Brustle, Vini Gautam, Ian Cockburn, Cathy Gillespie, Katharina Gaus, Woei Ming Lee

Proceedings Volume 10013, SPIE BioPhotonics Australasia; 100130K (2016) https://doi.org/10.1117/12.2242951
Event: SPIE BioPhotonics Australasia, 2016, Adelaide, Australia

Abstract

Intravital multiphoton microscopy has emerged as a powerful technique to visualize cellular processes in-vivo. Real time processes revealed through live imaging provided many opportunities to capture cellular activities in living animals. The typical parameters that determine the performance of multiphoton microscopy are speed, field of view, 3D imaging and imaging depth; many of these are important to achieving data from in-vivo. Here, we provide a full exposition of the flexible polygon mirror based high speed laser scanning multiphoton imaging system, PCI-6110 card (National Instruments) and high speed analog frame grabber card (Matrox Solios eA/XA), which allows for rapid adjustments between frame rates i.e. 5 Hz to 50 Hz with 512 × 512 pixels. Furthermore, a motion correction algorithm is also used to mitigate motion artifacts. A customized control software called Pscan 1.0 is developed for the system. This is then followed by calibration of the imaging performance of the system and a series of quantitative in-vitro and in-vivo imaging in neuronal tissues and mice.

Citation Download Citation

Yongxiao Li, Anne Brustle, Vini Gautam, Ian Cockburn, Cathy Gillespie, Katharina Gaus, and Woei Ming Lee "High speed multiphoton imaging", Proc. SPIE 10013, SPIE BioPhotonics Australasia, 100130K (9 December 2016); https://doi.org/10.1117/12.2242951

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