Huang, H.-C.H.-C.HuangChiang, S.-J.S.-J.ChiangWen, S.-H.S.-H.WenLee, P.-J.P.-J.LeeHUEI-WEN CHENChen, Y.-F.Y.-F.ChenCHEN-YUAN DONGYANG-FANG CHEN2022-03-092022-03-0920191083-3668https://www.scopus.com/inward/record.uri?eid=2-s2.0-85071574750&doi=10.1117%2f1.JBO.24.8.080502&partnerID=40&md5=31eba3afe961f0012ff88285a42a1a5dhttps://scholars.lib.ntu.edu.tw/handle/123456789/596818We acquired multiphoton images of normal and lung adenocarcinoma cell lines in three dimensions. Image stacks of the cells were then processed to obtain nucleus-to-cytoplasm (N/C) ratios in two and three dimensions. While N/C ratios in three dimensions can be unambiguously determined from the volumetric ratios of the nucleus and cytoplasm, two-dimensional (2-D) N/C can vary depending on the axial plane selected for N/C ratio determination. We determined 2-D N/C ratios from three criteria: (1) axial position at which the nuclear area is the largest; (2) the largest 2-D N/C ratio value; and (3) axial position at the midpoint of nuclear axial position. We found that different definitions of 2-D N/C ratio will significantly affect its value. Furthermore, in general, larger variance was found in 2-D rather than three-dimensional (3-D) N/C ratios. Lack of ambiguity in definition and reduced variance suggest that 3-D N/C ratio is a better parameter for characterizing tumor cells in the clinical setting. ? The Authors.[SDGs]SDG3[SDGs]SDG10[SDGs]SDG16Biological organs; Cell culture; Optics; Adenocarcinoma cells; Clinical settings; Lung Cancer; nucleus-to-cytoplasm -ratio; Three dimensions; Threedimensional (3-d); Two Dimensional (2 D); Two photon microscopy; Cells; cell nucleus; color; cytoplasm; diagnostic imaging; fluorescence microscopy; human; image processing; lung adenocarcinoma; lung tumor; metastasis; optical coherence tomography; procedures; three-dimensional imaging; tumor cell line; Adenocarcinoma of Lung; Cell Line, Tumor; Cell Nucleus; Color; Cytoplasm; Humans; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Lung Neoplasms; Microscopy, Fluorescence; Neoplasm Metastasis; Tomography, Optical CoherenceThree-dimensional nucleus-to-cytoplasm ratios provide better discrimination of normal and lung adenocarcinoma cells than in two dimensionsjournal article10.1117/1.JBO.24.8.0805022-s2.0-85071574750