COMBINED ATOMIC FORCE AND FLUORESCENCE MICROSCOPIES TO MEASURE SUBCELLULAR MECHANICAL PROPERTIES OF LIVE CELLS

Abstract

Atomic force microscopy (AFM) has been widely applied to study cellular functions;however, the relationship between cellular elasticity and ultrastructure density of a live cell remains to be discovered. The objective of this study was thus to extend our previous method of integrating AFM and immunofluorescence imaging to measure the ultrastructure distribution-related local mechanical properties of live cells. First, the morphology of a live cell was obtained by AFM. Second, the indentation sites were selected and flexible force volume indentation was performed. Third, the immunofluorescence image of the cell was obtained. The last was the mapping of the indentation site to the immunofluorescence image and obtaining the relationship between the local elastic properties and cytoskeleton density. The results on differentiated rat Schwann cells (RSCs) showed that the elastic modulus of stress fibers is higher than those of the nucleus and cytosol. The local elastic modulus of the live RSCs is correlated to the actin density, and the stress fiber that behaves like a pretension beam can give RSCs enough strength to envelop axons during myelination. In particular, the elastic properties of the live RSCs were twofold lower than those of the fixed. The results demonstrated the integrated method's applicability for a live cell.