A cell spheroid can be as large as 1 mm in diameter, containing more than 300,000 cells. It can be used for simulating a cancer tumor in cell treatment study. In this study, the cell uptake behaviors in cell spheroids are investigated. In particular, the penetration depths of Au nanoring, Au nanorod, and photosensitizer (AlPcS) in cell spheroids are evaluated based on a newly proposed technique of using confocal fluorescent microscopy. By using a geometric computation process, we can estimate the width of a fluorescent belt region, which corresponds to the uptake depth. Based on such evaluations, we can understand whether the quiescent viable cells in the middle layer of a cell spheroid. Also, by using a cell culture insert, we can estimate the uptake Au nanoparticle number per cell through mass spectrometry measurement. It is found that after 48-hour incubation time, the Au nanoparticles and photosensitizer all have the penetration depths in the range of 110-140 m. Such uptake penetration depths control the cell damage depths under laser illuminations. For observing the uptake depths, we use different fluorescent dyes to incubate the cell spheroids. It is found that the penetration depth of a dye also relies on its molecular weight. The smaller uptake penetration depths of Au nanoparticles into a cell spheroid can be attributed to the settlement of Au nanoparticles in the incubation solutions such that the upper portion of a cell spheroid has no access to the Au nanoparticles.
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