An improved technique for observing both membranous organelles and cytoskeleton in saponin-extracted cells.
Nobukazu Araki, T Lee, Hisashi Matsubara, Yutaka Takashima
Abstract
Nobukazu Araki, T Lee, Hisashi Matsubara, Yutaka Takashima
Abstract
Detergent-extraction of soluble cytoplasmic proteins under low osmotic pressure induces severe morphological damage, particularly on membranous organelles, although this processing is often required to observe cytoskeletal elements in the cells. In the present study, the osmotic adjustment of the rinsing solution and fixative, which were used after the detergent-extraction using saponin, has greatly improved the ultrastructure of membranous organelles. This improved method allowed us to observe both cytoskeletal elements and the fine structure of membranous organelles by electron microscopy.
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Detergent-extraction of soluble cytoplasmic proteins under low osmotic pressure induces severe morphological damage, particularly on membranous organelles, although this processing is often required to observe cytoskeletal elements in the cells. In the present study, the osmotic adjustment of the rinsing solution and fixative, which were used after the detergent-extraction using saponin, has greatly improved the ultrastructure of membranous organelles. This improved method allowed us to observe both cytoskeletal elements and the fine structure of membranous organelles by electron microscopy.
Key concepts: Organelle, Cytoskeleton, Fixative, Ultrastructure, Cytoplasm, Saponin, Electron microscope, Cell biology