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Cytological Analyses by Advanced Electron Microscopy

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Cyanidioschyzon merolae

Abstract

Recently several microscopy techniques have revealed the dynamics and structure of intracellular material such as organelles and supramolecular proteins for a better understanding of the fundamental properties of a cell. Electron microscopy provides information on extremely small biological specimens, providing images of several target molecules and organelles, including mitochondria and plastids, and their surrounding intracellular environments. Technical development using several advanced electron microscopes has been advanced to obtain 3D structural modeling. Among them, cryo-TEM and tomography techniques are useful for visualization of fine structures, but the observation area is limited. Conversely, “FIB-SEM” theoretically is not affected by the thickness of the sample, and FIB-SEM and reconstruction are effective for 3D structural modeling of C. merolae cells (2–5 microns) during cell division at the whole cell level. Another technique, UHVEM tomography, visualizes a detailed 3D structure of the light-harvesting complex on the surface of the thylakoid membrane in the plastid and the micro-compartments (cristae) in mitochondria. Furthermore, fine whole cell 3D structural model using cryo-STEM tomography, which is obtained under physical fixation and without staining, instead of chemical fixation, is compared with the models of FIB-SEM and 3D reconstruction.

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Abbreviations

3D:

Three-dimensional

ATUM-SEM:

Automated tape-collecting ultramicrotome

BSE:

Backscattered electron

CMOS:

Complementary metal oxide semiconductor

EM:

Electron microscopy

FIB-SEM:

Focused ion beam SEM

HAADF:

High-angle annular dark field

HPF:

High-pressure freezing

LM:

Light microscopy

SBF-SEM:

Serial block-face scanning electron microscopy

SBF-SEM:

Serial block-face SEM

SEM:

Scanning electron microscopy

STEM:

Scanning transmission electron microscopy

TEM:

Transmission electron microscopy

UHVEM:

Ultra-high-voltage electron microscopy

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Acknowledgments

We thank K. Ohta, Ph.D. (Kurume University) and T. Nishida, Ph.D. (Osaka University) for FIB-SEM operation/discussion and UHVEM operation, respectively.

Author information

Authors and Affiliations

  1. Cell Field Structure, Quantitative Biology Center, Riken, 3-10-23, Kagamiyama, 739-0046, Higashi-Hiroshima, Hiroshima, Japan

    Takako M. Ichinose & Atsuko H. Iwane

  2. Special Research Promotion Group, Graduate School of Frontier Biosciences, Osaka University, 1-3, Yamadaoka, 565-0871, Suita, Osaka, Japan

    Takako M. Ichinose & Atsuko H. Iwane

Authors
  1. Takako M. Ichinose
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  2. Atsuko H. Iwane
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Corresponding author

Correspondence to Atsuko H. Iwane .

Editor information

Editors and Affiliations

  1. Faculty of Science Department of Chemical and Biological Sciences, Japan Women’s University, Tokyo, Japan

    Tsuneyoshi Kuroiwa

  2. Department of Cell Genetics, National Institute of Genetics, Shizuoka, Japan

    Shinya Miyagishima

  3. Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Chiba, Japan

    Sachihiro Matsunaga

  4. Department of Life Sciences Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan

    Naoki Sato

  5. Department of Biological Sciences Graduate School of Science, University of Tokyo, Tokyo, Japan

    Hisayoshi Nozaki

  6. Laboratory for Chemistry and Life Science Institute of Innovative Research, Tokyo Institute of Technology, Kanagawa, Japan

    Kan Tanaka

  7. Graduate School of Science and Technology for Innovation Faculty of Science, Department of Biological Science and Chemistry, Yamaguchi University, Yamaguchi, Japan

    Osami Misumi

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Ichinose, T.M., Iwane, A.H. (2017). Cytological Analyses by Advanced Electron Microscopy. In: Kuroiwa, T., et al. Cyanidioschyzon merolae. Springer, Singapore. https://doi.org/10.1007/978-981-10-6101-1_9

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