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Recent Advances in Microwave-Assisted Specimen Processing for in-Situ Hybridization

Published online by Cambridge University Press:  02 July 2020

Mark A. Sanders  and
David M. Gartner
Mark A. Sanders
Affiliation:
Imaging Center, University of Minnesota, St., Paul, MN55108
David M. Gartner
Affiliation:
Imaging Center, University of Minnesota, St., Paul, MN55108
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Abstract

Co-localization of proteins and nucleic acid sequences by in situ hybridization and immunohistochemistry is frequently difficult as the procedure necessary to detect the target structure of one technique may negatively affect the target of the other. Structural impairments may also limit the application of the two techniques. to overcome these problems we developed a method to perform in situ hybridization and immunohistochemistry on Chromosome spreads, intact cells and semi-thin sections of methyl methacrylate-embedded tissue. Microwavestimulated antigen retrieval, signal amplification by catalyzed reporter deposition, and fluorescent dyes were used for all techniques, yielding high sensitivity and excellent morphological preservation compared to conventional processing techniques. Co-localization of in situ hybridization and immunohistochemistry probes with high morphological resolution was achieved using computerized image reconstruction. The latter may allow for the co-localization of multiple antigens and a specific DNA sequence at the same tissue level.

Type
Recent Techniques for the Fixation and Staining of Biological Samples (Organized by M. Sanders and K. McDonald)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 1198 - 1199
Copyright
Copyright © Microscopy Society of America 2001

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References

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