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Localization-based super-resolution imaging meets high-content screening
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DOI：10.1038/NMETH.4486.png)
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Abstract
En 中文
Single-molecule localization microscopy techniques have proven to be essential tools for quantitatively monitoring biological processes at unprecedented spatial resolution. However, these techniques are very low throughput and are not yet compatible with fully automated, multiparametric cellular assays. This shortcoming is primarily due to the huge amount of data generated during imaging and the lack of software for automation and dedicated data mining. We describe an automated quantitative single-molecule-based super-resolution methodology that operates in standard multiwell plates and uses analysis based on high-content screening and datamining software. The workflow is compatible with fixed-and live-cell imaging and allows extraction of quantitative data like fluorophore photophysics, protein clustering or dynamic behavior of biomolecules. We demonstrate that the method is compatible with high-content screening using 3D dSTORM and DN A-PAINT based super-resolution microscopy as well as single-particle tracking.
Keywords:
OPTICAL RECONSTRUCTION MICROSCOPY
HIGH-DENSITY
HIGH-THROUGHPUT
FLUORESCENT-PROBES
AMPA RECEPTORS
LIVING CELLS
DNA-PAINT
MOLECULE
PROTEINS
FLUOROPHORES
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