Department of Embryology, Carnegie Institution for Science, Baltimore, MD 21218, USA.
Methods. 2010 May;51(1):45-51. doi: 10.1016/j.ymeth.2009.12.010. Epub 2010 Jan 6.
One can manually isolate the giant oocyte nucleus or germinal vesicle (GV) of Xenopus from a living oocyte with nothing more complicated than jewelers' forceps and a dissecting microscope. Similarly, one can remove the nuclear envelope by hand and allow the lampbrush chromosomes and other nuclear organelles to spread on a microscope slide. After centrifugation, the nuclear contents adhere tightly to the slide, where they can be subjected to immunostaining or fluorescent in situ hybridization for visualization by conventional or confocal microscopy. Preparations of isolated GV contents reveal details of nuclear structure that are almost impossible to attain by more conventional techniques.
人们可以用珠宝镊子和解剖显微镜从活体卵母细胞中手动分离爪蟾的巨大卵母细胞核或生发泡(GV)。同样,人们可以用手去除核膜,并让灯刷染色体和其他核细胞器在显微镜载玻片上展开。离心后,核内容物紧密附着在载玻片上,在那里可以进行免疫染色或荧光原位杂交,通过传统或共聚焦显微镜进行可视化。分离的 GV 内容物的制备揭示了核结构的细节,这几乎是传统技术无法达到的。