Mechanical stimulation of individual stereocilia of living cochlear hair cells by atomic force microscopy

• Published in: Ultramicroscopy 2000-02, Vol.82 (1), p.269-278
• Main Authors: Langer, M.G, Koitschev, A, Haase, H, Rexhausen, U, Hörber, J.K.H, Ruppersberg, J.P
• Format: Article
• Language: English
• Subjects: AFM; Animals; Atomic force microscopy; Cilia - physiology; Elasticity; Fixatives; Hair cells; Hair Cells, Auditory, Inner - physiology; Hair Cells, Auditory, Inner - ultrastructure; Hair Cells, Auditory, Outer - physiology; Hair Cells, Auditory, Outer - ultrastructure; Medical imaging; Microscopy, Atomic Force - instrumentation; Microscopy, Atomic Force - methods; Microscopy, Electron, Scanning - methods; Morphology; Physical Stimulation; Rats; Scanning electron microscopy; Stereocilia; Stiffness
• ISSN: 0304-3991; 1879-2723
• DOI: 10.1016/S0304-3991(99)00136-9

This paper describes the investigation of elastical properties and imaging of living cochlear hair bundles of inner (IHC) and outer hair cells (OHC) on the level of individual stereocilia. A custom-made AFM-setup was used, allowing to scan the mechano-sensitive structures of the inner ear under direct control of an upright differential interference contrast (DIC) microscope with a water-immersion objective. Scanning electron microscopy (SEM) images of the identical hair bundles obtained after AFM investigation demonstrated that forces up to 1.5 nanonewton (nN) did not cause obvious damage of the surface morphology of the stereocilia. These are the first images of hair bundles of living sensory cells of the organ of Corti by AFM. They display the tips of individual stereocilia and the typical V-shape of ciliary bundles. Since line scans clearly show that slope and force interaction depend on the elastical properties of stereocilia, quantitative stiffness measurements and stimulation of single transduction channels are suggested.