Non-invasive single-cell biomechanical analysis using live-imaging datasets

J Cell Sci. 2016 Sep 1;129(17):3351-64. doi: 10.1242/jcs.191205. Epub 2016 Jul 15.

Abstract

The physiological state of a cell is governed by a multitude of processes and can be described by a combination of mechanical, spatial and temporal properties. Quantifying cell dynamics at multiple scales is essential for comprehensive studies of cellular function, and remains a challenge for traditional end-point assays. We introduce an efficient, non-invasive computational tool that takes time-lapse images as input to automatically detect, segment and analyze unlabeled live cells; the program then outputs kinematic cellular shape and migration parameters, while simultaneously measuring cellular stiffness and viscosity. We demonstrate the capabilities of the program by testing it on human mesenchymal stem cells (huMSCs) induced to differentiate towards the osteoblastic (huOB) lineage, and T-lymphocyte cells (T cells) of naïve and stimulated phenotypes. The program detected relative cellular stiffness differences in huMSCs and huOBs that were comparable to those obtained with studies that utilize atomic force microscopy; it further distinguished naïve from stimulated T cells, based on characteristics necessary to invoke an immune response. In summary, we introduce an integrated tool to decipher spatiotemporal and intracellular dynamics of cells, providing a new and alternative approach for cell characterization.

Keywords: Cell biomechanics; Cell migration; Live imaging; Mechanobiology; Mesenchymal stem cells; T-lymphocyte cells.

MeSH terms

  • Adult
  • Algorithms
  • Animals
  • Automation
  • Biomechanical Phenomena / drug effects
  • CD8-Positive T-Lymphocytes / cytology
  • CD8-Positive T-Lymphocytes / drug effects
  • Cell Survival / drug effects
  • Databases as Topic*
  • Elasticity
  • Humans
  • Imaging, Three-Dimensional*
  • Ionomycin / pharmacology
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Mice, Inbred C57BL
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Phenotype
  • Single-Cell Analysis / methods*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors
  • Viscosity

Substances

  • Ionomycin
  • Tetradecanoylphorbol Acetate

Associated data

  • figshare/figshare.com/s/a207830096d7cdd32164