期刊
BIOTECHNIQUES
卷 55, 期 5, 页码 245-+出版社
BIOTECHNIQUES OFFICE
DOI: 10.2144/000114099
关键词
super-resolution microscopy; dSTORM; high-content screening; correlative microscopy; Golgi complex
资金
- Methoden fur die Lebenswissenschaften of the Baden-Wurttemberg Stiftung [P-LS-SPII/11]
- New methods in systems biology program of the Federal Ministry of Education and Research [0315523A]
- CellNetworksCluster of Excellence [EXC81]
High-throughput microscopy is an effective tool for rapidly collecting data on a large scale. However, high throughput comes at the cost of low spatial resolution. Here we introduce correlative light microscopy by combining fast automated widefield imaging, confocal microscopy and super-resolution microscopy. We demonstrate the potential of this approach for scalable experiments. The workflow consists of a robust approach for selecting cells of interest on a wide-field screening microscope at low resolution and subsequently re-localizing those cells with micrometer precision for confocal and super-resolution imaging. As a case study, we visualized and quantified cis- and trans-Golgi markers at increasing resolution. Method Summary: Quantitative description of biological systems is the major challenge of biological research. Fluorescence microscopy plays an important role because it allows researchers to not only visualize but also evaluate virtually every cellular process of interest. Fluorescence microscopy, however, comes with a trade-off between throughput and spatial/temporal resolution. Due to these technological limitations, imaging biological samples is generally constrained to either high throughput but low resolution or high resolution but low throughput.
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