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一种基于显微镜的方法,用于研究活的和固定的人类卵母细胞中的减数分裂。

A microscopy-based approach for studying meiosis in live and fixed human oocytes.

作者信息

Zielinska Agata P, Schuh Melina

机构信息

Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

Max Planck Institute for Biophysical Chemistry, Department of Meiosis, Göttingen, Germany.

出版信息

Methods Cell Biol. 2018;145:315-333. doi: 10.1016/bs.mcb.2018.03.039. Epub 2018 May 26.

Abstract

Human eggs frequently carry an incorrect number of chromosomes, which is a leading cause of pregnancy loss and congenital disorders. The origins of high aneuploidy rates in human eggs have remained largely unclear. This is due to two main reasons: first, the availability of human eggs is limited so that studies of fixed human eggs typically involve very small numbers and limited quantifications. Second, methods for studying meiosis in live human eggs have been missing. The ever rising prevalence of Assisted Reproductive Technologies has facilitated a recent breakthrough in the field. The mechanistic basis of meiosis in humans can now be examined directly in live eggs. Here, we present a robust method for culturing human eggs in vitro and describe how meiotic processes in human eggs can be studied in real time using fluorescent reporters. We further describe methods for the in-depth analysis of immunolabeled eggs by super-resolution light microscopy.

摘要

人类卵子常常携带数目不正确的染色体,这是导致妊娠丢失和先天性疾病的主要原因。人类卵子中高非整倍体率的起源在很大程度上仍不清楚。这主要有两个原因:第一,人类卵子的可获得性有限,因此对固定的人类卵子的研究通常涉及数量非常少且定量有限。第二,一直缺少在活的人类卵子中研究减数分裂的方法。辅助生殖技术日益增长的普及率促成了该领域最近的一项突破。现在可以直接在活卵子中检查人类减数分裂的机制基础。在此,我们提出一种体外培养人类卵子的可靠方法,并描述如何使用荧光报告分子实时研究人类卵子中的减数分裂过程。我们还进一步描述了通过超分辨率光学显微镜对免疫标记卵子进行深入分析的方法。

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