Abstract
Chromatin organization influences gene and transposon expression, and regulates various cellular processes. Higher order chromatin structure has been widely studied using genomic approaches and microscopy image analyses. Chromosome conformation capture and sequencing the junction of DNA fragments enables the study of both chromatin interaction and chromosome folding. However, certain cell types are embedded in other cell types which complicate the process of studying them using high-throughput genomic approaches. To overcome this limitation, high-resolution microscopy techniques are now available to investigate chromatin organization in single cells. In this chapter, we provide a detailed protocol to prepare chromosome spreading from tomato nuclei, to label genomic loci by fluorescence in situ hybridization, and to visualize these locations at high resolution with Structured Illumination microscopy.
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Acknowledgments
We would like to thank Alessandro di Maio (University of Birmingham) for his advice and knowledge of SIM. This work was supported by a BBSRC grant-aided support as part of the Institute Strategic Programme designing Future Wheat Grant (BP/P016855/1) and the BBSRC grant 1644151.
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Kuo, P., Darbyshire, A., Lambing, C. (2022). Super-resolution Chromatin Visualization Using a Combined Method of Fluorescence In Situ Hybridization and Structured Illumination Microscopy in Solanum lycopersicum. In: Lambing, C. (eds) Plant Gametogenesis. Methods in Molecular Biology, vol 2484. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2253-7_7
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DOI: https://doi.org/10.1007/978-1-0716-2253-7_7
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Publisher Name: Humana, New York, NY
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