Patel Snehal, Novikova Natalya, Beenders Brent, Austin Christopher, Bellini Michel
Department of Biochemistry and College of Medicine, School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Chromosome Res. 2008;16(2):223-32. doi: 10.1007/s10577-007-1189-z. Epub 2008 Jan 10.
The nucleus of an amphibian oocyte can be manually isolated in mineral oil where it maintains all its activities for several hours. These undisrupted (live) nuclei have been used successfully in recent years to analyze the dynamic organization of several non-chromosomal nuclear organelles. Here, we describe an improved procedure for imaging an oil-isolated nucleus by light microscopy and we use it to produce the very first images of lampbrush chromosomes in an in vivo-like condition. These chromosomes are morphologically identical to those observed in conventional nuclear spread preparations. Importantly, their lateral loops, which are active RNA polymerase II transcription units, are readily distinguished by differential interference contrast microscopy.
两栖类卵母细胞的细胞核可以在矿物油中手动分离出来,在这种情况下它能维持数小时的所有活动。近年来,这些未受破坏(活的)细胞核已成功用于分析几种非染色体核细胞器的动态组织。在这里,我们描述了一种通过光学显微镜对油分离细胞核进行成像的改进方法,并利用它首次在类似体内的条件下获得灯刷染色体的图像。这些染色体在形态上与传统核铺展制备中观察到的染色体相同。重要的是,它们的侧环是活跃的RNA聚合酶II转录单位,通过微分干涉对比显微镜很容易区分。