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基于显微镜的温度敏感 S. pombe 细胞活力定量分析方案。

Microscopy-based protocol for the quantification of cells viability for temperature-sensitive S. pombe.

机构信息

Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.

Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore; National University Health System, Singapore, Singapore; NUS Center for Cancer Research, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore; NUS Graduate School-Integrative Sciences & Engineering Programme, National University of Singapore, Singapore, Singapore.

出版信息

STAR Protoc. 2024 Jun 21;5(2):103076. doi: 10.1016/j.xpro.2024.103076. Epub 2024 May 19.

DOI:10.1016/j.xpro.2024.103076
PMID:38768030
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11111821/
Abstract

Conventional colony-forming unit assay to measure cell viability is laborious and results in large experimental variability, which prohibits accurate quantification of microbial viability. Here, we present a microscopy-based protocol for the quantification of cells viability for temperature-sensitive S. pombe. We describe steps for growing and treating yeast cells and visualization of individual cells viability based on Phloxine B staining. We then detail procedures for data processing using Nikon NIS Elements Advanced Research (AR) software. For complete details on the use and execution of this protocol, please refer to Lim et al..

摘要

传统的集落形成单位检测法用于测量细胞活力既繁琐，又会导致较大的实验变异性，从而无法准确量化微生物的活力。在此，我们提供了一种基于显微镜的方法来检测温度敏感型 S. pombe 的细胞活力。我们描述了培养和处理酵母细胞的步骤，并基于 Phloxine B 染色来观察单个细胞的活力。然后，我们详细介绍了使用 Nikon NIS Elements Advanced Research (AR) 软件进行数据处理的过程。有关此协议的使用和执行的完整详细信息，请参阅 Lim 等人的文章。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/46176709a032/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/ffce6120a826/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/6496507dfa11/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/ada47c3a7f5f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/0418f27a9ba6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/cf50d0490e1d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/46176709a032/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/ffce6120a826/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/6496507dfa11/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/ada47c3a7f5f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/0418f27a9ba6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/cf50d0490e1d/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2963/11111821/46176709a032/gr5.jpg

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