Advances in imaging the interphase nucleus using thin cryosections
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The mammalian genome is partitioned amongst various chromosomes and encodes for approximately 30,000 protein-coding genes. Gene expression occurs after exit from mitosis, when chromosomes partially decondense within the cell nucleus to allow the enzymatic activities that work on chromatin to access each gene in a regulated fashion. Differential patterns of gene expression evolve during cell differentiation to give rise to the over 200 cell types in higher eukaryotes. The architectural organisation of the genome inside the interphase cell nucleus, and associated enzymatic activities, reveals dynamic and functional compartmentalization of the genome. In this review, I highlight the advantages of Tokuyasu cryosectioning on the investigation of nuclear structure and function.
KeywordsNuclear compartments Gene expression Chromosome territories Transcription factories Confocal microscopy Correlative microscopy
I would like to thank David J. Vaux for first suggesting the use of Tokuyasu cryosections, Mike Hollinshead for generously sharing his knowledge about cryosectioning and for many fruitful collaborations, my laboratory co-workers, especially Sonya Martin, Sheila Q. Xie, and Miguel R. Branco, for their contribution in furthering the development of high-resolution labelling strategies, Sheila Q. Xie and Emily Brookes for comments on the manuscript, and Miguel R. Branco for the kind gift of images in Figs. 2a and 3, and Sheila Xie for image 1b. The work in my laboratory is supported by the Medical Research Council (UK).
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