Solovei Irina
Institute of Human Genetics, Biozentrum (LMU), University of Münich, Planegg-Martinsried, Germany.
Methods Mol Biol. 2010;659:71-82. doi: 10.1007/978-1-60761-789-1_5.
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.
在对核功能空间组织的理解方面，近期的进展很大程度上归功于荧光原位杂交（FISH）技术。使用该技术的大多数研究都是在培养细胞中进行的。然而，全生物体中的核过程在很大程度上可能与培养细胞中的不同，并且实际上在不同组织之间也不相似。因此，为了更好地理解离体生物体材料中的核过程，有必要研究组织切片中的核组织。由于方法学问题，切片上的FISH在核生物学研究中仍然不常见。本章提出的方案基于多年来在各种组织的冷冻切片上杂交不同探针的经验。
