酵母
活体细胞成像
免疫荧光
细胞生物学
细胞
间接免疫荧光
生物
细胞壁
化学
抗体
生物化学
免疫学
出处
期刊:Methods in molecular biology
日期:2014-01-01
卷期号:: 79-90
被引量:25
标识
DOI:10.1007/978-1-4939-1363-3_6
摘要
In spite of their small size, the cellular morphology, structure, and protein localization of yeast cells can be successfully imaged. A detailed protocol for preparing yeast cells for live-cell imaging is described, including techniques to immobilize yeast for time-lapse microscopy. Protocols for indirect immunofluorescence are outlined, including strategies for fixation, cell wall digestion, and the use of primary and secondary antibodies conjugated to fluorescent moieties. Alternative approaches to these techniques are discussed, highlighting the advantages and disadvantages where possible. Using these protocols, investigation of yeast cell structure and protein localization will continue to yield important insights into yeast cell biology and regulation.
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