Nanoscale Imaging of Primary Cilia with Scanning Ion Conductance Microscopy

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Abstract
Primary cilia are hair-like sensory organelles whose dimensions and location vary with cell type and culture condition. Herein, we employed scanning ion conductance microscopy (SICM) to visualize the topography of primary cilia from different cell types. By combining SICM with fluorescence imaging, we successfully distinguished between surface cilia that project outward from the cell surface and subsurface cilia that are trapped below it. The nanoscale structure of the ciliary pocket, which cannot be easily identified using a confocal fluorescence microscope, was observed in SICM images. Furthermore, we developed a topographic reconstruction method using current-distance profiles to evaluate the relationship between set point and topographic image and found that a low set point is important for detecting the true topography of a primary cilium using hopping mode SICM.
Funding Information
  • Development of Advanced Measurement and Analysis Systems
  • Takeda Science Foundation
  • Precursory Research for Embryonic Science and Technology
  • Japan Agency for Medical Research and Development
  • Japan Society for the Promotion of Science (15H05422, 15K20856)
  • Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering

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