FISH on 3D preserved bovine and murine preimplantation embryos

Fluorescence in situ hybridization (FISH) is a commonly used technique for the visualization of whole chromosomes or subchromosomal regions, such as chromosome arms, bands, centromeres, or single gene loci. FISH is routinely performed on chromosome spreads, as well as on three-dimensionally preserve...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 659; p. 437
Main Authors Koehler, Daniela, Zakhartchenko, Valeri, Ketterl, Nina, Wolf, Eckhard, Cremer, Thomas, Brero, Alessandro
Format Journal Article
LanguageEnglish
Published United States 2010
Subjects
Animals
Blastocyst - cytology
Blastocyst - metabolism
Cattle
Imaging, Three-Dimensional
In Situ Hybridization, Fluorescence - methods
Mice
Microscopy, Fluorescence
Nucleic Acid Denaturation
Tissue Embedding
Tissue Fixation
Tissue Preservation - methods
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Summary:Fluorescence in situ hybridization (FISH) is a commonly used technique for the visualization of whole chromosomes or subchromosomal regions, such as chromosome arms, bands, centromeres, or single gene loci. FISH is routinely performed on chromosome spreads, as well as on three-dimensionally preserved cells or tissues (3D FISH). We have developed 3D FISH protocol for mammalian preimplantation embryos to investigate the nuclear organization of chromosome territories and subchromosomal regions during the first developmental stages. In contrast to cells, embryos have much more depth and their nuclei are therefore less accessible to probes used to visualize specific genomic regions by FISH. The present protocol was developed to establish a balance between sufficient embryo permeabilization and maximum preservation of nuclear morphology.
ISSN:1940-6029
DOI:10.1007/978-1-60761-789-1_34