期刊
METHODS AND APPLICATIONS IN FLUORESCENCE
卷 3, 期 3, 页码 -出版社
IOP Publishing Ltd
DOI: 10.1088/2050-6120/3/3/034006
关键词
fluorescence microscopy; super-resolution; single molecule localization; direct stochastic optical reconstruction; multicolor; registration; single molecule noise
资金
- German Funding Agency (DFG) [SFB958/Z02]
- Marie Curie International Reintegration Grant [256501]
Multicolor single molecule localization-based super-resolution microscopy (SMLM) approaches are challenged by channel crosstalk and errors in multi-channel registration. We recently introduced a spectral demixing-based variant of direct stochastic optical reconstruction microscopy (SDdSTORM) to perform multicolor SMLM with minimal color crosstalk. Here, we demonstrate that the spectral demixing procedure is inherently free of errors in multicolor registration and therefore does not require multicolor channel alignment. Furthermore, spectral demixing significantly reduces single molecule noise and is applicable to astigmatism-based 3D multicolor imaging achieving 25 nm lateral and 66 nm axial resolution on cellular nanostructures.
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