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Single-Molecule Fluorescence Imaging in Living Saccharomyces cerevisiae Cells.

ABSTRACT: This protocol describes how to image fluorescently tagged proteins, RNA, or DNA inside living Saccharomyces cerevisiae cells at the single-molecule level. Imaging inside living cells, as opposed to fixed materials, gives access to real-time kinetic information. Although various single-molecule imaging applications are discussed, we focus on imaging of gene transcription at the single-RNA level. To obtain the best possible results, it is important that both imaging parameters and yeast culture conditions are optimized. Here, both aspects are described. For complete details on the use and execution of this protocol, please refer to Lenstra et al. (2015) and Donovan et al. (2019).

SUBMITTER: Brouwer I

PROVIDER: S-EPMC7757289 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Single-Molecule Fluorescence Imaging in Living <i>Saccharomyces cerevisiae</i> Cells.

Brouwer Ineke I Patel Heta Piyush HP Meeussen Joseph Victor Willem JVW Pomp Wim W Lenstra Tineke Laura TL

STAR protocols 20201017 3

This protocol describes how to image fluorescently tagged proteins, RNA, or DNA inside living <i>Saccharomyces cerevisiae</i> cells at the single-molecule level. Imaging inside living cells, as opposed to fixed materials, gives access to real-time kinetic information. Although various single-molecule imaging applications are discussed, we focus on imaging of gene transcription at the single-RNA level. To obtain the best possible results, it is important that both imaging parameters and yeast cul ...[more]

PMID: 33377036

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