Development of an in-solution observation method using atmospheric scanning electron microscopy (ASEM)

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Other Title
  • 大気圧電子顕微鏡ASEMによる水中観察法の開発
  • ―半導体の超薄膜技術とバイオ顕微鏡の融合研究―
  • — Interdisciplinary research between semiconductor fabrication technology and biological electron microscopy —

Description

Protein complexes in cells and tissues play critical roles in various physiological functions, including embryogenesis and signal processing.To observe the dynamics of protein complexes, high-resolution and high-throughput electron microscopy (EM) in aqueous solution is required. However, standard EM requires the sample to be in a vacuum. With ASEM, an inverted scanning electron microscope (SEM) observes the wet sample from beneath an open dish while an optical microscope (OM) observes it from above. The disposable dish with a silicon nitride film window can hold a few milliliters of culture medium, allowing various types of cells to be cultured in a stable environment. This system was used to develop in situ correlative OM/SEM immuno-microscopy in liquid. Using this method, we have observed a dynamic string-like gathering of STIM1 on the endoplasmic reticulum in Jurkat T cells in response to Ca2+ store depletion. We have also observed filamentous-actin (F-actin) and tubulin in the growth cones of primary-culture neurons, as well as in synapses. Further,we have monitored in-situ electrochemical reactions in electrolytes, and the melting and solidification of solder using ASEM.

Journal

  • Synthesiology English edition

    Synthesiology English edition 8 (4), 162-173, 2015

    National Institute of Advanced Industrial Science and Technology

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