Dual-wavelength photoacoustic molecular microscopy for three-dimensional cell culture systems

• Main Authors: Po-Yi Lee, 李柏逸
• Other Authors: 李百祺
• Format: Others
• Language: zh-TW
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/51378926256752952454

碩士 === 國立臺灣大學 === 生醫電子與資訊學研究所 === 105 === With the knowledge of cell interaction between tumor and therapeutic cells, it can help to develop cell-based therapeutic strategies such as immunotherapy. In contrast to traditional two-dimensional (2D) cell cultures, a three-dimensional (3D) cell culture system better mimics the cell environment for observation of cellular processes and understanding biological mechanisms. However, conventional molecular imaging methods such as confocal microscopy and two-photon microscopy hardly depict cell processes in 3D models. Thus, we have developed dual-wavelength photoacoustic molecular microscopy for 3D cell culture systems for visualizing cell distribution and morphology. We employed gold nanoparticles (AuNPs) as wavelength-dependent contrast agents in our dual-wavelength optical resolution photoacoustic microscope, with the wavelength of 523 nm and 800 nm, for visualizing CD8+ cytotoxic T lymphocytes (CTLs) in an in vitro 3D tumor microenvironment. Targeted CTLs with avidin-conjugated gold nanospheres (AuNSs) and hepatocellular carcinoma labeled with gold nanorods (AuNRs) were distinguishable by their distinct absorption spectra under 523-nm and 800-nm laser irradiation. We successfully showed that dual-wavelength OR-PAM can map the distribution of CTLs with AuNSs under 523-nm irradiation and the 3D morphology of tumorspheres with AuNRs under 800-nm irradiation in an in vitro 3D microenvironment.