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Multicolor super\u2010resolution 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\u2010parameter detection strategy, which uses both the fluorescence activation wavelength and the emission color to discriminate between photo\u2010activatable fluorescent probes. First, we obtained two\u2010color super\u2010resolution images using the near\u2010infrared 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\u2010activation enabled multi\u2010parameter detection of six different probes. Using this approach, we obtained six\u2010color super\u2010resolution 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\u2010diffraction\u2010limit resolution.<\/jats:p>","DOI":"10.1002\/cphc.201100735","type":"journal-article","created":{"date-parts":[[2011,12,24]],"date-time":"2011-12-24T16:21:41Z","timestamp":1324743701000},"page":"99-107","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":149,"title":["Multicolor Super\u2010Resolution Fluorescence Imaging via Multi\u2010Parameter Fluorophore Detection"],"prefix":"10.1002","volume":"13","author":[{"given":"Mark","family":"Bates","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Graham T.","family":"Dempsey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kok 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