The nucleus is the largest organelle in the cell. When deformed with techniques like Atomic Force Microscopy or micropipette aspiration, the nucleus appears to be elastic and much stiffer than the cytoplasm. Whether the nucleus behaves like a stiff elastic object when shaped by cellular forces and on physiological time scales, such as during migration through confining channels, is not clear. Here I will discuss our efforts to understand nuclear mechanics in cell migration. I will present live cell imaging experiments that reveal surprising nuclear mechanical behaviors such as drop-like deformation. I will show how the nucleus is likely shaped by viscous coupling between the nucleus and the cytoplasm rather than static cytoskeletal stresses. I will conclude with an example of high content imaging of cancer nuclear morphology for drug discovery applications.
Recent advances in photodynamic inactivation and photobiomodulation require extensive research of application safety in living tissues in vitro and in vivo. Superficial phototoxicity induced cellular morphological changes have been observed and recorded with using confocal Brillouin microspectrometer. We are reporting evidence of biomechanical processes occurring in cells subjected to high-power laser radiation. 4T1 murine fibroblast cells were used in the study, making results easily after exposure to high power laser radiation. Spatial distribution of subcellular structures’ stiffness was recorded with high precision and analyzed, drawing correlation between existing morphological model and novel stiffness data within the cell.
Cancer remains among the leading causes of death in the United States. Early detection, classification and understanding of malignant cell proliferation and metastasis mechanisms are crucial for effective treatment. Current malignant cell studies largely rely on either invasive imaging techniques or invasive research protocols that hinder both speed and accuracy of cancer research. Here we are reporting successful imaging of cancer metastasis processes on a cellular level using Brillouin microspectroscopic imaging. In this research we are specifically presenting results of a non-invasive interrogation of elastic properties of 4T1 murine fibroblast cells in a spheroid model acquired with our custom-built confocal Brillouin microspectrometer. Spatial map of elastic properties was recorded for both interior and exterior regions of the 4T1 cell spheroid. We observed lower stiffness of cancer cells compared to cells from internal regions. In addition we observed the difference in stiffness values between cells exposed to challenging and normal environmental conditions. Our findings correlate well with prior published data, acquired with conventional biomechanical assessment techniques.
The nucleus is the largest organelle in the cell. When deformed with techniques like AFM or micropipette aspiration, the nucleus appears to be highly elastic and much stiffer than the cytoplasm. Whether the nucleus behaves like a stiff elastic object when shaped by cellular forces and on physiological time scales, such as during migration through confining channels, is not clear. Here I will discuss our efforts to understand nuclear mechanics in cell migration. I will present live cell imaging experiments that reveal surprising nuclear mechanical behaviors such as drop-like deformation. I will show how the nucleus is likely shaped by viscous coupling between the nucleus and the cytoplasm rather than static cytoskeletal stresses.
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