Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-05T05:29:52.827Z Has data issue: false hasContentIssue false
  • English
  • Français

Integrating Light and TEM Information with F-TEM images

Published online by Cambridge University Press:  14 March 2018

P.A. Sims ,
C.A. Lockwood  and
J.D. Hardin
P.A. Sims*
Affiliation:
Zoology Department, University of Wisconsin
C.A. Lockwood
Affiliation:
Zoology Department, University of Wisconsin
J.D. Hardin
Affiliation:
Zoology Department, University of Wisconsin
*
[email protected]

Extract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Fluorescent fusion proteins are widely used to visualize the localization of proteins in worms, fish, flies and tissue culture cells. We have used two different methods that use high pressure freezing (HPF) combined with correlative light microscopy (LM) and TEM. The first method uses fluorescence from live organisms immobilized in agarose followed by HPF and standard freeze substitution in dry solvent with osmium. This pre-embedding method optimizes ultrastructural preservation. A second, post-embedding method preserves fluorescence and immunoreactivity from embedded and polymerized thin sections. Here we describe post-embedding fluorescent integrated TEM images (F-TEM).

Type
Research Article
Information
Microscopy Today , Volume 13 , Issue 5 , September 2005 , pp. 16 - 19
Copyright
Copyright © Microscopy Society of America 2005

References

[1] Robinson, JM, et. al.J. Histo Cyto. 49(7) (2001) 803–808 CrossRefGoogle Scholar
[2] Takizawa, T. and Robinson, J. J. Histo Cyto. 51(6) (2003) 707–714.CrossRefGoogle Scholar
[3] Luby-Phelps, et. al. J. Histo Cyto. 51(3)(2003) 271–274.CrossRefGoogle Scholar
[4] Inoue, S (1995). In Pawley, JB ed. Handbook of Biol. Confocal Microscopy.Google Scholar
[5](Heim, R., Tsien, R. and Zuker, C., unpublished) in Cubitt et. al. TIBS 20 Nov. 1995.Google Scholar
[6] Webster, , Paul Microscopy Today November 2000 (00-9): 28-34CrossRefGoogle Scholar
[7] Griffiths, G. 1993. Fine Structure Immunocytochemistry. Springer Verlag, Heidelberg & Berlin.CrossRefGoogle Scholar
[8] Phillippe, Rostaing et. al, J. Histo Cyto. 52(1)(2004) 1–12.Google Scholar
[9] Walther, P., Ziegler, A., J. of Microscopy 208, Ptl. Oct 2002.CrossRefGoogle Scholar
[10] Ward, W.W. (1988) in Green Fluorescnet Protein: Proteins, Applications and Protocols (Chalfie, M. and Kain, S., Eds.) 1st ed., pp.45–75, Wiley, New York.Google Scholar
[11] Campbell, R. E., (PNAS) vol. 99(12) (2002) 7877–7882.CrossRefGoogle Scholar
[12] Koppen, et. al. Nature Cell Biol 3 (2001) 983–991 Google Scholar