Cryogenic Cooling of Biological Samples for Electron and Optical Microscopy

  • G. J. Fuld
  • R. G. Hansen
  • N. Loes
Part of the Applications of Cryogenic Technology book series (APCT, volume 10)


Cryogenic liquid transfer systems have proven to be the method of choice for most cooling applications for spectrophotometric studies. The application of such methods to biological systems requires a few modifications. The cryogenic systems can operate in any position but most liquid samples must be placed in cuvettes in the vertical position from the top. Temperature control from 77K to room temperature can be accomplished using a liquid nitrogen supply, a 20 ohm strip heater, a silicon diode transducer and an automatic temperature controller.

Various designs are presented with limited test data, where standard cuvettes are mounted on OHFC copper in a vacuum shroud necessary for the low temperature studies. Some suggested approaches to expanding the versatility of the systems are given. A unique application is the use of a vacuum cooled sample plate and injecting the test sample into the chamber through a rubber septum giving instantaneous cooling of the sample.


Radiation Shield Cryogenic Cool Cold Finger Liquid Cryogen OFHC Copper 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    B. Meyer, “Low Temperature Spectroscopy” American Elsevier Publishing, New York (1971).Google Scholar
  2. 2.
    F. Franks, “Biophysics and Biochemistry at Low Temperatures,” Cambridge University Press, Cambridge (1985)Google Scholar
  3. 3.
    A. W. Robards, and U. B. Sleytr, Low Temperature Methods in Biological Electron Microscopy in “Practical Methods in Electron Microscopy,” Vol. 10, Am M. Glauert, ed., Elsevier Press, Amsterdam (1985)Google Scholar
  4. 4.
    R. G. Hansen & Associates, Private Communication and “High-Tran Liquid Transfer System,” data sheets, 1988.Google Scholar
  5. 5.
    Units supplied by Scientific Instruments, West Palm Beach, FL, or LakeShore Cryotronics, Westerville, OH, are generally applicable.Google Scholar
  6. 6.
    SEM stage cryogenic devices are offered by Oxford Instrument Co., Oxford, England, and R. G. Hansen & Associates, Santa Barbara, CA.Google Scholar
  7. 7.
    J. Bastacky, C. Goodman, and T. L. Hayes, A Specimen Holder for Low-Temperature Scanning Electron Microscopy, J. Electron Microscopy Technique,. 14: 83–84 (1990).CrossRefGoogle Scholar
  8. 8.
    S. J. Shah, K. R. Diller, and S. J. Aggarwal, A Personal Computer-Based Temperature Control System for Cryomicroscopy, Cryobiology, 24: 163–167 (1987).PubMedCrossRefGoogle Scholar
  9. 9.
    J. A. Sargent, Temperature Scanning Electron Microscopy: Advantages and Applications, Scanning Microscopy, 2: 835–849 (1988).PubMedGoogle Scholar
  10. 10.
    K. R. Diller, Cryomicroscopy in “Low Temperature Biotechnology Emerging Applications and Engineering Contributions,” J. J. McGrath and K. R. Diller, eds. ASME, BED Vol 10, HTD Vol 98, New York (1989).Google Scholar
  11. 11.
    P. L. Steponkus, M. F. Dowert, J. R. Ferguson and R. L. Levin, Cryomicroscopy of isolated plant protoplasts, Cryobiology, 21: 209–233 (1984).CrossRefGoogle Scholar
  12. 12.
    Ch. Korber, S. Englich, P. Schwindke, M. W. Scheiwe, G. Rau, A. Hubel, and E. G. Cravalho Low temperature light microscopy and its application to study of freezing in aqueous solutions and biological cell applications, J. Microscopy, 141: 263–276 (1985).CrossRefGoogle Scholar
  13. 13.
    R. G. Hansen & Associates, “Liquid Cryogen Pourfill Dewar,” Bulletin PFD 189.Google Scholar
  14. 14.
    S. Badulescu, R. Bicca De Alencastro, H. LeThanh, G. Richer, C. Sandorfy, P-P. Vaudreuil, and D. Vocelle, The Protonation of a Retinyl Schiff Base: A Study by FTIR Spectroscopy at Low Temperature in Solution, Photochemistry and Photobiology, 49:313–318(1989).Google Scholar
  15. 15.
    S. R. Harder, B. A. Feinberg, and S. W. Ragsdale, A Spectroelectrochemical Cell Designed for Low Temperature Electron Paramagnetic Resonance Titration of Oxygen-Sensitive Proteins, Analytical Biochemistry, 181: 283–287 (1989).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • G. J. Fuld
    • 1
  • R. G. Hansen
    • 1
  • N. Loes
    • 1
  1. 1.R. G. Hansen and AssociatesSanta BarbaraUSA

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