생물학분야에서 Cryo-electron Tomography 활용기법

Techniques for Cryo-electron Tomography in Biological Field

  • 문지영 (고려대학교 생명과학대학 생명공학원 세포 및 생체 3차 구조 연구실) ;
  • 이경은 (고려대학교 생명과학대학 생명공학원 세포 및 생체 3차 구조 연구실) ;
  • 한성식 (고려대학교 생명과학대학 생명공학원 세포 및 생체 3차 구조 연구실)
  • Mun, Ji-Young (Laboratory of Cell Engineering & 3-D Structure, Graduate School of Life Sciences and Biotechnology, Korea University) ;
  • Lee, Kyung-Eun (Laboratory of Cell Engineering & 3-D Structure, Graduate School of Life Sciences and Biotechnology, Korea University) ;
  • Han, Sung-Sik (Laboratory of Cell Engineering & 3-D Structure, Graduate School of Life Sciences and Biotechnology, Korea University)
  • 발행 : 2008.06.30

초록

현재 생물학 분야에서 세포 구조를 3차로 구현하기 위해 전자 현미경을 이용하는 연구는 매우 빠르게 발전하고 있다. 최근에는 시료의 구조를 변형시키는 화학 고정법 대신 시료를 빠른 시간 내에 동결고정시킴으로써 구조 변형 없이 세포 3차 구조 구현이 가능해졌다. 이러한 기술의 도입으로 단편이미지로만 세포 구조를 이해해왔던 지금까지와 달리 Cryo-ET를 통해 인위적인 구조 변형 없는 3차 구조 구현이 가능해졌고, 이 기법은 약물전달, 나노 공학 등 여러 학문 분야에 활용됨으로써 학문 발전에 이바지 할 것이다.

In Biology, Studies Using Electron Microscopy for making Cell Structure to 3D reconstruction very fast development. Recently, by using Cryo fixation, we can see cell 3D structure without structural change, instead of using chemical fixation which can change cell structure. Before using this technology, we could understand cell structures only in 2D images. But now, through cryo-ET, 3D reconstruction of cell structure without artificial structure changes can be possible and this technology will give us many advantages in Drug delivery and Nanothechnology.

키워드

참고문헌

  1. Al-Amoudia A, Norlenb LPO, Dubochet J: Cryo-electron microscopy of vitreous sections of native biological cells and tissues. J Struct Biol 148 : 131-135, 2004 https://doi.org/10.1016/j.jsb.2004.03.010
  2. Almgren M, Edwards K, Karlsson Gr: Cryo transmission electron microscopy of liposomes and related structures. Colloids Surf A Physicochem Eng Asp 174 : 3-21, 2000 https://doi.org/10.1016/S0927-7757(00)00516-1
  3. Bohm J, Frangakis AS, Hegerl R, Nickell S, Typke D, Baumeister W: Toward detecting and identifying macromolecules in a cellular context: Template matching applied to electron tomograms. PNAS 97 : 14245-14250, 2000 https://doi.org/10.1073/pnas.230282097
  4. Ding B, Turgeon R, Parthasarathy MV: Routine cryofixation of plant tissue by propane jet freezing for freeze substitution. J Electron Microsc Tech 19 : 107-117, 1991 https://doi.org/10.1002/jemt.1060190111
  5. Duman JG, Pathak NJ, Ladinsky MS, McDonald KL, Forte JG: Three-dimensional reconstruction of cytoplasmic membrane networks in parietal cells. J Cell Sci 115 : 1251-1258, 2002
  6. Hess MW: Of plants and other pets:practical aspects of freezesubstitution and resin embedding. J Microsc 212 : 44-52, 2003 https://doi.org/10.1046/j.1365-2818.2003.01237.x
  7. Kellenberger E: The response of biological macromolecules and supramolecular structures to physics of specimen cryopreparation. In:Steinbrecht RA, Zeirold K, ed, Cryotechniques in biological electron microscopy. pp. 35-63, Springer-Verlag, Berlin Heidelberg, 1987
  8. Lee KE, Han SS: Cryo-elctron tomography. Mol Cell Biol news 19 : 26-35, 2007
  9. Lucic V, Forster F, Baumeister W: Structural studies by electron tomography: from cells to molecules. Annu Rev Biochem 74 : 833-865, 2005 https://doi.org/10.1146/annurev.biochem.73.011303.074112
  10. McDonald K: New developments and applications of high pressure freezing. Microsc Microanal 11 : 1092-1093, 2005
  11. Monaghan P, Perusinghe N, Muller M: High-pressure freezing for immunocytochemistry. J Microsc 192 : 248-258, 1998 https://doi.org/10.1046/j.1365-2818.1998.00387.x
  12. Nicastro D, McIntosh R, Baumeister W: 3D structure of eukaryotic flagella in a quiescent state revealed by cryo-electron tomography. PNAS 102 : 15889-15894, 2005 https://doi.org/10.1073/pnas.0508274102
  13. Reipert S, Fischer I, Wiche G: High-pressure freezing of epithelial cells on sapphire coverslips. J Microsc 21 : 81-85, 2004
  14. Robards AW, Wilson AJ: Low-temperature Methods for TEM and SEM. In: Robards AW, Wilson AJ, eds, Procedures in electron microscopy, pp. 16:11.11-16:18.18, John Wiley & Sons, New York, 1997