Zebrafish are an attractive animal model for the study of disease due to their low cost, ease of care, and ability to rapidly produce large colonies. Some mutant zebrafish, for example those from the casper line, exhibit optical properties which make them ideal specimens for in vivo optical imaging. In this study we use an ultra-high frequency photoacoustic (PA) microscope to image the head and trunk of live casper fish, and generate 3D renderings of the local functional vasculature with single-cell resolution.
Five day-post-fertilization larvae were anesthetized using tricane and embedded in 1.5% (w/v) agarose to prevent movement during image acquisition. A transmission mode PA microscope equipped with a single element transducer (either 200 or 400 MHz central frequency) and a 532 nm laser focused through a 4X optical objective was used to raster scan the larvae. Reconstructions of the vasculature were created from the recorded RF data.
Images of the trunk showed the caudal artery (CA) and vein (CV), as well as intersegmental vessels (ISV). The ISVs were observed to originate from either the CA or CV, and curve around the notochord to join at the dorsal longitudinal anastomotic vessel. Furthermore, individual erythrocytes were resolved within the caudal hematopoietic tissue. Reconstructions of the head revealed a tortuous organization of developing vessels in the brain. This work is the first demonstration of in vivo PAM at frequencies greater than 200 MHz, and paves the way for future studies which will explore label-free imaging of nanoparticles and cancer progression in vivo.
Michael J. Moore, Suzan El-Rass, Yongliang Xiao, Xiao-Yan Wen, and Michael C. Kolios, "Ultra-high frequency photoacoustic microscopy of zebrafish larvae in vivo (Conference Presentation)," Proc. SPIE 10494, Photons Plus Ultrasound: Imaging and Sensing 2018, 104942O (Presented at SPIE BiOS: January 31, 2018; Published: 15 March 2018); https://doi.org/10.1117/12.2291181.5752222402001.
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