Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence <em>In Situ</em> Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice

Rupak Pathak; Igor Koturbash; Martin Hauer-Jensen

doi:10.3791/55162

通过多色荧光跨染色体畸变稳定的检测原位杂交（渔业部）和光谱核型分析（SKY）的照射小鼠

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Genetics

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JoVE Journal Genetics

Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice

通过多色荧光跨染色体畸变稳定的检测原位杂交（渔业部）和光谱核型分析（SKY）的照射小鼠

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10:14 min

January 11, 2017

DOI: 10.3791/55162-v

Rupak Pathak¹, Igor Koturbash², Martin Hauer-Jensen^1,3

¹Division of Radiation Health, Department of Pharmaceutical Sciences, College of Pharmacy,University of Arkansas for Medical Sciences, ²Department of Environmental Health, Fay W. Boozman School of Public Health,University of Arkansas for Medical Sciences, ³Surgical Service,Central Arkansas Veterans Healthcare System

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本方案描述了多重荧光原位杂交（mFISH）和光谱核型（SKY）在识别小鼠全身照射后骨髓细胞染色体间稳定畸变的有用性。

这种分子细胞生成方法的总体目标是观察暴露于全身照射的小鼠骨髓细胞中的染色体间稳定畸变。这种方法可以帮助回答放射生物学领域的关键问题，例如暴露于全身辐射的小鼠骨髓细胞中诱导稳定染色体畸变的风险。该技术的主要优点是它允许对辐射诱导的小鼠骨髓细胞中稳定的遗传损伤进行引导可视化，这些损伤可以通过许多细胞代传播。

从小鼠股骨和胫骨中分离骨髓并根据文本方案制备单细胞悬液后，小心地将细胞悬液覆盖在等体积的骨髓单核细胞分离培养基上。在室温下以 400 x G 离心梯度 30 分钟。然后，小心地收集血沉棕黄层，不要干扰其余层，并将溶液转移到新的 15 毫升离心管中。

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