ALGAE

  • 제21권4호
  • /
  • Pages.479-483
  • /
  • 2006
  • /
  • 1226-2617(pISSN)
  • /
  • 2093-0860(eISSN)
한국조류학회(藻類) (The Korean Society of Phycology)
권호 표지
DOI QR코드

DOI QR Code

High Pressure Freezing (HPF)을 이용한 조류 Ptilota filicina의 미세구조 관찰:HPF 고정법과 화학 고정법의 비교

Ultrastructures of Ptilota filicina (Rhodophyta) by High Pressure Freezing(HPF): Comparison of HPF Fixation and Chemical Fixation

  • 이상희 (한국기초과학지원연구원 전자현미경연구부) ;
  • 김윤중 (한국기초과학지원연구원 전자현미경연구부) ;
  • 정종만 (한국기초과학지원연구원 전자현미경연구부) ;
  • 김진규 (한국기초과학지원연구원 전자현미경연구부) ;
  • 김영민 (한국기초과학지원연구원 전자현미경연구부) ;
  • 권희석 (한국기초과학지원연구원 전자현미경연구부) ;
  • 문원진 (한국기초과학지원연구원 광주센터) ;
  • 이석훈 (한국기초과학지원연구원 전자현미경연구부)
  • Lee, Sang-Hee (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Kim, Youn-Joong (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Jeong, Jong-Man (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Kim, Jin-Gyu (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Kim, Young-Min (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Kweon, Hee-Seok (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Moon, Won-Jin (KBSI, Gwang-ju Center) ;
  • Lee, Seok-Hoon (Division of Electron Microscopic Research, Korea Basic Science Institute)
  • 발행 : 2006.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

In preparation of the biological samples for electron microscopy, the chemical fixation by glutaraldehyde, paraformaldehyde, and OsO4 has been generally used for a long time. However, the chemical fixation method has some problems: the infiltration time is a little bit long and the ultrastructure of cell or tissue transforms before complete fixation of sample. So, recently, cryo-fixation is considered more often in biomedical field. In this study, we compared High Pressure Freezing (HPF) method with chemical fixation method using a algal sample (Ptilota filicina J. Agardh), which was difficult to fix using chemical fixation method. In chloroplast, the ultrastructure of thylakoid lamella and phycobilisome can not show clearly by chemical fixation. In this study we could observe the ultrastructure of thylakoid lamella and phycobilisome of chloroplast very clearly using HPF fixation. An improved images of ultrastructures of nucleus, mitochondrion and floridean starch could obtain. These results suggest that HPF method is very useful method in algal specimen for electron microscopy.

키워드

참고문헌

  1. Aichinger N. and Lutz-Meindl U. 2005. Organelle interactions and possible degradation pathways visualized in highpressure frozen algal cells. J. Microsc. 219: 86-94 https://doi.org/10.1111/j.1365-2818.2005.01496.x
  2. Ding R., McDonald K.L. and McIntosh J.R. 1993. Threedimensional reconstruction and analysis of mitotic spindles from the yeast Schizzosaccharomyces pombe. J. Cell Biol. 120:141-151 https://doi.org/10.1083/jcb.120.1.141
  3. Echlin P. 1992. Low-Temperature Microscopy and Analysis. Plenum Press, New York and London
  4. Erk I., Nicolas G., Caroff A. and Lepault J. 1998. Electron microscopy of frozen biological objects: a study using cryosectioning and cryosubstitution. J. Microsc. 189: 236-248 https://doi.org/10.1046/j.1365-2818.1998.00323.x
  5. Hohenberg H., Mannweiler K. and Muller M. 1994. Highpressure freezing of cell suspensions in cellulose capillary tubes. J. Microsc. 175: 34-43 https://doi.org/10.1111/j.1365-2818.1994.tb04785.x
  6. Moor H. 1987. Theory and practice of high pressure freezing. In: R. Steinbrecht and K. Zierold (eds), Cryotechniques in Biological Electron Microscopy. Springer-Verlag, Berlin. pp. 175-191
  7. Reynolds E.S. 1963. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17: 208-212 https://doi.org/10.1083/jcb.17.1.208
  8. Riehle U. and Hochli M. 1973. The theory and technique of high-pressure freezing. In: E. L. Benedetti and P. Favard (eds), Freeze-Etching Technique and Application. Societe Francaise de microscopie Electronique, Paris. pp. 31-60
  9. Shimoni E. and Muller M. 1998. On optimizing high-pressure freezing: from heat transfer theory to a new microbiopsy device. J. Microsc. 192: 236-247 https://doi.org/10.1046/j.1365-2818.1998.00389.x
  10. Spurr A.R. 1969. A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26: 31-43 https://doi.org/10.1016/S0022-5320(69)90033-1
  11. Studer D., Michel M., Wohlwend M., Hunziker E.B. and Buschmann M.D. 1995. Vitrification of articular cartilage by high-pressure freezing. J. Microsc. 179: 321-332 https://doi.org/10.1111/j.1365-2818.1995.tb03648.x
  12. Thijssen M.H., Van Went J.L. and Van Aelst A.C. 1998. Heptane and isooctane as embedding fluids for high-pressure freezing of Petunia ovules followed by freeze-substitution. J. Microsc. 192: 228-235 https://doi.org/10.1046/j.1365-2818.1998.00385.x
  13. Walther P., Muller M. and Schweingruber M.E. 1984. The ultrastructure of the cell surface and plasma membrane of exponential and stationary phase cells of Schizosaccharomyces pombe, grown in different media. Arch. Microbiol. 137: 128-134 https://doi.org/10.1007/BF00414453
  14. Walther P., Schweingruber A.M. Muller M. and Schweingruber M.E. 1988. Morphological organization of glycoprotein containing cell surface structures in yeast. J. Ultrastruct. Molec. Struct. Res. 101: 123-136 https://doi.org/10.1016/0889-1605(88)90002-X
  15. Walther P. and Ziegler A. 2002. Freeze substitution of highpressure frozen samples: the visibility of biological membranes is improved when the substitution medium contains water. J. Microsc. 208: 3-10 https://doi.org/10.1046/j.1365-2818.2002.01064.x

피인용 문헌

  1. Ascophyllumand its symbionts. X. Ultrastructure of the interaction betweenA. nodosum(Phaeophyceae) andMycophycias ascophylli(Ascomycetes) vol.86, pp.2, 2008, https://doi.org/10.1139/B07-122