For emerging tissue-engineering applications, transplants, and cell-based therapies it is important to assess cell viability and function in vivo in deep tissues. Bioluminescence and fluorescence methods are poorly suited to deep monitoring applications with high resolution and require genetically-engineered reporters which are not always feasible. We report on a method for imaging cell viability using deep, high-resolution photoacoustic imaging. We use an exogenous dye, Resazurin, itself weakly fluorescent until it is reduced from blue to a pink color with bright red fluorescence. Upon cell death fluorescence is lost and an absorption shift is observed. The irreversible reaction of resazurin to resorufin is proportional to aerobic respiration. We detect colorimetric absorption shifts using multispectral photoacoustic imaging and quantify the fraction of viable cells. SKOV-3 cells with and without ±80oC heat treatment were imaged after Resazurin treatment. High 575nm:620nm ratiometric absorption and photoacoustic signals in viable cells were observed with a much lower ratio in low-viability populations.
Roger J. Zemp and Robert J. Paproski, "Multimodality molecular imaging and extracellular vesicle release based genetic profiling with porphyrin nanodroplets (Conference Presentation)," Proc. SPIE 10064, Photons Plus Ultrasound: Imaging and Sensing 2017, 100642F (Presented at SPIE BiOS: February 01, 2017; Published: 24 April 2017); https://doi.org/10.1117/12.2253254.5379284522001.
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Study of self-shadowing effect as a simple means to realize nanostructured thin films and layers with special attentions to birefringent obliquely deposited thin films and photo-luminescent porous silicon