Abstract
Recent progress in the understanding of the spatial organization of nuclear functions owes a lot to fluorescence in situ hybridization (FISH) methodology. The majority of studies using this technology have been carried out using cultured cells. However, nuclear processes in whole organisms, may be to a notable degree, different from those in cultured cells and actually not similar across different tissues. Therefore, for better understanding of nuclear processes in ex vivo organismal material, it is necessary to study nuclear organization in sections of tissue. FISH on sections is still not common in nuclear biology studies mostly due to methodological problems. The protocol suggested in this chapter is based on several years experience in hybridizing different probes on cryosections of various tissues.
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References
Cremer, M., Grasser, F., Lanctot, C., Müller, S., Neußer, M., Zinner, R., Solovei, I., and Cremer, T. (2008) Multicolor 3D Fluorescence In Situ Hybridization for Imaging Interphase Chromosomes, in Methods in Molecular Biology: The Nucleus (Hancock, R., Ed.) pp 205–239, Humana Press.
Cremer, M., Müller, S., Köhler, D., Brero, A., and Solovei, I. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4723.
Solovei, I., and Cremer, M. (2010) Combination of Immunostaining and 3D-FISH on Cultured Cells, in Methods in Molecular Biology: FISH (Bridger J. M., V. E., Ed.) Humana Press.
Solovei, I., Walter, J., Cremer, M., Habermann, H., Schermelleh, L., and Cremer, T. (2002) FISH on Three-Dimensionally Preserved Nuclei, in FISH (Beatty B., Larson S. M., J. Squire, Ed.) pp 119–157, Oxford University Press, Oxford.
Solovei, I., Grasser, F., and Lanctôt, C. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4729.
Shi, S. R., Cote, R. J., and Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: past, present, and future. J Histochem Cytochem 45, 327–43.
Shi, S. R., Cote, R. J., and Taylor, C. R. (2001) Antigen retrieval techniques: current perspectives. J Histochem Cytochem 49, 931–7.
Henegariu, O., Bray-Ward, P., and Ward, D. C. (2000) Custom fluorescent-nucleotide synthesis as an alternative method for nucleic acid labeling. Nat Biotechnol 18, 345–8.
Müller, S., Neusser, M., Köhler, D., and Cremer, M. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4730.
Nimmakayalu, M., Henegariu, O., Ward, D. C., and Bray-Ward, P. (2000) Simple method for preparation of fluor/hapten-labeled dUTP. Biotechniques 28, 518–22.
Acknowledgments
The author thanks Yana Feodorova and Süleyman Kösem who, as graduate students, carried out some of the immuno-FISH experiments described in this protocol and Mike Fessing (University of Bradford) for providing BAC DNA for Rps27. This work was supported by DFG grant SO1054/1.
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Solovei, I. (2010). Fluorescence in situ Hybridization (FISH) on Tissue Cryosections. In: Bridger, J., Volpi, E. (eds) Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology, vol 659. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-789-1_5
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DOI: https://doi.org/10.1007/978-1-60761-789-1_5
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