Microwave fixation of Setaria Digitata for scanning electron microscopy

선충류의 주사전자현미경적 관찰을 위한 마이크로웨이브 고정법

  • Lee, San-Soo (Biotherapy Human Resources Center, College of Veterinary Medicine, Chonnam National University) ;
  • Cho, Kyoung-Oh (Biotherapy Human Resources Center, College of Veterinary Medicine, Chonnam National University) ;
  • Shin, Kil-Sang (College of Natural Sciences, Soonchunhyang University) ;
  • Shin, Sung-Shik (Biotherapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
  • Accepted : 2007.05.14
  • Published : 2007.06.30

Abstract

Conventional processing of biological materials including nematode parasites for scanning electron microscopy includes fixation with glutaraldehyde and osmium, followed by dehydration in an ascending grade of ethanol, and finally freeze drying. This procedure takes about 8 to 12 h depending on the characteristics of samples. Microwave irradiation of 2,450 MHz enhance the action of cross-linking fixatives and can greatly accelerate various stages of tissue processing. In this study, samples of nematode parasites, Setaria digitata, were fixed by a combination of conventional chemical fixation and the microwave irradiation during the process. The microwave irradiation was also incorporated in the serial dehydration process with ethanol. The complete procedure from the initial fixation to the completion of dehydration with ethanol was reduced to 1 h with good preservation of the ultrastructural details of the specimens.

References

  1. 신길상. 적외선 온도감응기를 장착한 마이크로파 오븐의 생체물질 고정효과. Korean J Zool 1994, 37, 144-145
  2. Amako K, Umeda A. Bacterial surfaces as revealed by the high resolution scanning electron microscope. J Gen Microbiol 1977, 98, 297-299 https://doi.org/10.1099/00221287-98-1-297
  3. Argall K, Armati P. Use of microwave fixation in the preparation of cell cultures for observation with the scanning electron microscope. J Electron Microsc Tech 1990, 16, 347-350 https://doi.org/10.1002/jemt.1060160406
  4. Carranza GA, Cruz D, Wagner GG. Microwave fixation:in situ tick (Acari: Ixodidae) histoanatomy, thin sectioning of tick tissues, and antigen preservation in mouse spleen. J Med Entomol 1990, 27, 1067-1070 https://doi.org/10.1093/jmedent/27.6.1067
  5. Dawes CJ. Biological Techniques for Transmission and Scanning Electron Microscopy. pp. 241-252, Ladd Research Industries, Burlington, 1984
  6. Epperlein HH, Schwarz H, Piendl T, Lofberg J, Studer D, Spring H, Mller M. Improved preservation of subepidermal extracellular matrix in axolotl embryos using electron microscopical techniques based on cryoimmobilization. J Struct Biol 1997, 118, 43-61 https://doi.org/10.1006/jsbi.1996.3838
  7. Fox NE, Demaree RS. Quick bacterial microwave fixation technique for scanning electron microscopy. Microsc Res Tech 1999, 46, 338-339 https://doi.org/10.1002/(SICI)1097-0029(19990815/01)46:4/5<338::AID-JEMT12>3.0.CO;2-L
  8. Giberson RT, Demaree RS. Microwave fixation:understanding the variables to achieve rapid reproducible results. Microsc Res Tech 1995, 32, 246-254 https://doi.org/10.1002/jemt.1070320307
  9. Hernandez F, Guillen R. Microwave processing fo scanning electron microscopy. Eur J Morphol 2000, 38, 109-111 https://doi.org/10.1076/0924-3860(200004)38:2;1-F;FT109
  10. Hopwood D, Milne G, Penston J. A comparison of microwaves and heat alone in the preparation of tissue for electron microscopy. Histochem J 1990, 22, 358-364 https://doi.org/10.1007/BF01003170
  11. Horobin RW. Problems and artifacts of microwave accelerated procedures in neurohistotechnology and resolutions. Methods 1998, 15, 101-106 https://doi.org/10.1006/meth.1998.0612
  12. Ibsch M, Vohringer P, Anken RH, Rahmann H. Electronmicroscopic investigations on the role of vesiclelike bodies in inner maculae for fish otolith growth. Adv Space Res 2000, 25, 2031-2034 https://doi.org/10.1016/S0273-1177(99)01011-X
  13. Innes JR, Shoho C. Cerebrospinal nematodiasis: focal encephalomyelomalacia of animals caused by nematodes (Setaria digitata); a disease which may occur in man. AMA Arch Neurol Psychiatry 1953, 70, 325-349 https://doi.org/10.1001/archneurpsyc.1953.02320330050006
  14. InoueT, Osatake H. A new drying method of biological specimens for scanning electron microscopy: the t-butyl alcohol freeze-drying method. Arch Histol Cytol 1988, 51, 53-59 https://doi.org/10.1679/aohc.51.53
  15. Kaab MJ, Richards RG. Chondrocytes in articular cartilage preserved by rapid microwave enhanced fixation. Bone 1995, 17, 557-596
  16. Koga D, Ueno M, Yamashina S. SEM observation of the insect prepared by microwave irradiation. J Electron Microsc (Tokyo) 2003, 52, 477-484 https://doi.org/10.1093/jmicro/52.5.477
  17. Login GR, Dvorak AM. Methods of microwave fixation for microscopy. A review of research and clinical applications: 1970-1992. Prog Histochem Cytochem 1994, 27, 1-127
  18. Login GR. Microwave fixation versus formalin fixation of surgical and autopsy tissue. Am J Med Technol 1978, 44, 435-437
  19. Login GR, Ku TC, Dvorak AM. Rapid primary microwave-aldehyde and microwave-osmium fixation: Improved detection of rat parotid acinar cell secretory granule $\alpha$-amylase using a post-embedding immunogold ultrastructural morphometric analysis. J histochem Cytochem 1995, 43, 515-523 https://doi.org/10.1177/43.5.7730590
  20. Login GR, Quissel DO, Dvorak AM. Immunocytochemical detection of amylase in microwave fixed parotid acinar cells. J Dent Res 1992, 71, 304
  21. Mayers CP. Histological fixation by microwave heating. J Clin Pathol 1970, 23, 273-275 https://doi.org/10.1136/jcp.23.3.273
  22. Mizuhira V, Hasegawa H, Notoya M. Microwave fixation and localization of calcium in synaptic vesicles. J Neurosci Methods 1994, 55, 125-136 https://doi.org/10.1016/0165-0270(94)90205-4
  23. Rhee JK, Choi EY, Park BK, Jang BG. Application of scanning electron microscopy in assessing the prevalence of some Setaria species in Korean cattle. Korean J Parasitol 1994, 32, 1-6 https://doi.org/10.3347/kjp.1994.32.1.1
  24. Smid HM, Schooneveld H, Meerloo T. Microwave fixation of water-cooled insect tissues for immunohistochemistry. Histochem J 1990, 22, 313-320 https://doi.org/10.1007/BF01003163