In Situ Mechanical Characterization of the Cell Nucleus by Atomic Force Microscopy

ACS Nano. 2014 Apr 22;8(4):3821-8. doi: 10.1021/nn500553z. Epub 2014 Apr 1.

Abstract

The study of nuclear mechanical properties can provide insights into nuclear dynamics and its role in cellular mechanotransduction. While several methods have been developed to characterize nuclear mechanical properties, direct intracellular probing of the nucleus in situ is challenging. Here, a modified AFM (atomic force microscopy) needle penetration technique is demonstrated to mechanically characterize cell nuclei in situ. Cytoplasmic and nuclear stiffness were determined based on two different segments on the AFM indentation curves and were correlated with simultaneous confocal Z-stack microscopy reconstructions. On the basis of direct intracellular measurement, we show that the isolated nuclei from fibroblast-like cells exhibited significantly lower Young's moduli than intact nuclei in situ. We also show that there is in situ nucleus softening in the highly metastatic bladder cancer cell line T24 when compared to its less metastatic counterpart RT4. This technique has potential to become a reliable quantitative measurement tool for intracellular mechanics studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomechanical Phenomena
  • Cell Line
  • Cell Nucleus / metabolism*
  • Fibroblasts / cytology
  • Humans
  • Image Processing, Computer-Assisted
  • Mechanical Phenomena*
  • Mechanotransduction, Cellular
  • Microscopy, Atomic Force / instrumentation
  • Microscopy, Atomic Force / methods*
  • Needles