MICROSCOPY Vol.45▶No.2 2010
■Feature Articles: Autophagy: Fusion of Morphology and Molecular Biology

Artificial Induction of Autophagy around Polystyrene Microbeads

Shouhei Kobayashi and Tokuko Haraguchi

Abstract: Autophagy is a dynamic cellular process, which can take place in indefinite places within the cytoplasm, making it difficult to predict when and where it occurs. To overcome such difficulties, we have developed a new method to induce autophagy in defined places around polystyrene beads internalized by non-phagocytotic cells, and also developed a method of live cell fluorescence imaging combined with electron microscopy, designated “live CLEM” (correlative light and electron microscopy after live imaging). This technology provides a unique opportunity to visualize dynamic processes of autophagy within the cell in the living state and in high resolution. In this paper, we demonstrate live CLEM imaging of the beads internalized into living cells, and discuss how this method is powerful to understand cellular processes associated with autophagy.

Key words: fluorescence microscopy,electron microscopy, Correlative light and electron microscopy after live cell imaging, polystyrene bead, autophagy