DOI: 10.1002/cphc.201100735. Multicolor Super‐Resolution Fluorescence Imaging via Multi‐Parameter Fluorophore Detection

Multicolor Super‐Resolution Fluorescence Imaging via Multi‐Parameter Fluorophore Detection

10.1002/cphc.201100735

Crossref journal-article

Wiley

ChemPhysChem (311)

Abstract

AbstractUnderstanding the complexity of the cellular environment will benefit from the ability to unambiguously resolve multiple cellular components, simultaneously and with nanometer‐scale spatial resolution. Multicolor super‐resolution fluorescence microscopy techniques have been developed to achieve this goal, yet challenges remain in terms of the number of targets that can be simultaneously imaged and the crosstalk between color channels. Herein, we demonstrate multicolor stochastic optical reconstruction microscopy (STORM) based on a multi‐parameter detection strategy, which uses both the fluorescence activation wavelength and the emission color to discriminate between photo‐activatable fluorescent probes. First, we obtained two‐color super‐resolution images using the near‐infrared cyanine dye Alexa 750 in conjunction with a red cyanine dye Alexa 647, and quantified color crosstalk levels and image registration accuracy. Combinatorial pairing of these two switchable dyes with fluorophores which enhance photo‐activation enabled multi‐parameter detection of six different probes. Using this approach, we obtained six‐color super‐resolution fluorescence images of a model sample. The combination of multiple fluorescence detection parameters for improved fluorophore discrimination promises to substantially enhance our ability to visualize multiple cellular targets with sub‐diffraction‐limit resolution.

Bibliography

Bates, M., Dempsey, G. T., Chen, K. H., & Zhuang, X. (2011). Multicolor SuperâResolution Fluorescence Imaging via MultiâParameter Fluorophore Detection. ChemPhysChem, 13(1), 99â107. Portico.

Authors 4

Mark Bates (first)
Graham T. Dempsey (additional)
Kok Hao Chen (additional)
Xiaowei Zhuang (additional)

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Dates

Type	When
Created	13 years, 8 months ago (Dec. 24, 2011, 4:21 p.m.)
Deposited	1 year, 10 months ago (Oct. 16, 2023, 11:05 a.m.)
Indexed	2 months, 1 week ago (June 23, 2025, 3:28 a.m.)
Issued	13 years, 8 months ago (Dec. 23, 2011)
Published	13 years, 8 months ago (Dec. 23, 2011)
Published Online	13 years, 8 months ago (Dec. 23, 2011)
Published Print	13 years, 7 months ago (Jan. 16, 2012)

Funders 0

None

BibTeX

@article{Bates_2011, title={Multicolor Super‐Resolution Fluorescence Imaging via Multi‐Parameter Fluorophore Detection}, volume={13}, ISSN={1439-7641}, url={http://dx.doi.org/10.1002/cphc.201100735}, DOI={10.1002/cphc.201100735}, number={1}, journal={ChemPhysChem}, publisher={Wiley}, author={Bates, Mark and Dempsey, Graham T. and Chen, Kok Hao and Zhuang, Xiaowei}, year={2011}, month=dec, pages={99–107} }

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