Quantitative analysis and comparison of 3D morphology between viable and apoptotic MCF-7 breast cancer cells and characterization of nuclear fragmentation.
Yuhua Wen, Zhan Chen, Jianfen Lu, Elizabeth Ables, Jean-Luc Scemama, Li V Yang, Jun Q Lu, Xin-Hua Hu
Author Information
Yuhua Wen: Institute for Advanced Optics, Hunan Institute of Science and Technology, Yueyang, Hunan, China.
Zhan Chen: Department of Physics, East Carolina University, Greenville, North Carolina, United States of America.
Jianfen Lu: Department of Biology, East Carolina University, Greenville, North Carolina, United States of America.
Elizabeth Ables: Department of Biology, East Carolina University, Greenville, North Carolina, United States of America.
Jean-Luc Scemama: Department of Biology, East Carolina University, Greenville, North Carolina, United States of America.
Li V Yang: Department of Internal Medicine, Brody School of Medicine, East Carolina University, Greenville, North Carolina, United States of America.
Jun Q Lu: Institute for Advanced Optics, Hunan Institute of Science and Technology, Yueyang, Hunan, China.
Xin-Hua Hu: Institute for Advanced Optics, Hunan Institute of Science and Technology, Yueyang, Hunan, China. ORCID
Morphological changes in apoptotic cells provide essential markers for defining and detection of apoptosis as a fundamental mechanism of cell death. Among these changes, the nuclear fragmentation and condensation have been regarded as the important markers but quantitative characterization of these changes is yet to be achieved. We have acquired confocal image stacks of 206 viable and apoptotic MCF-7 cells stained by three fluorescent dyes. Three-dimensional (3D) parameters were extracted to quantify and compare their differences in morphology. To analyze nuclear fragmentation, a new method has been developed to determine clustering of nuclear voxels in the reconstructed cells due to fluorescence intensity changes in nuclei of apoptotic cells. The results of these studies reveal that the 3D morphological changes in cytoplasm and nuclear membranes in apoptotic cells provide sensitive targets for label-free detection and staging of apoptosis. Furthermore, the clustering analysis and morphological data on nuclear fragmentation are highly useful for derivation of optical cell models and simulation of diffraction images to investigate light scattering by early apoptotic cells, which can lead to future development of label-free and rapid methods of apoptosis assay based on cell morphology.
