3D positioning of tagged DNA loci by widefield and super-resolution fluorescence imaging of fixed yeast nuclei

STAR Protocols

Philippe Pasero¹  Julien Cau¹  Julio Mateos-Langerak¹  Armelle Lengronne¹  Jérôme Poli¹  Mégane Da Mota¹ 

[1] Institut de Génétique Humaine, CNRS, Université de Montpellier, 34396 Montpellier, France;

关键词: Cell culture;    Single Cell;    Genetics;    Microscopy;    Model Organisms;    Gene Expression;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: This protocol describes how to culture, image, and determine the nuclear position of a fluorescently tagged DNA locus in the 3D nucleoplasm of fixed Saccharomyces cerevisiae cells. Here, we propose a manual scoring method based on widefield images and an automated method based on 3D-SIM images. Yeast culture conditions have to be followed meticulously to get the best biological response in a given environment.For complete details on the use and execution of this protocol, please refer to Forey et al. (2020).

【 授权许可】

Unknown