Nat. Methods 8, 253–259 (2011); published online 6 February 2011; corrected online 16 February 2011; corrected after print 30 August 2012; corrected after print 3 August 2015

In the version of this article initially published, the sequence reported for the dBrainbow construct was incorrect. The blue fluorescent protein was reported as EBFP2; it is mTFP1. The mKO2 protein is tagged with a V5 epitope and not a Myc epitope, and the EGFP protein is tagged with HSV, not with V5, as detailed in a previous correction to this paper. The red channel reflects endogenous mKO2 fluorescence, not anti-Myc staining. The mKO2 protein is produced by a codon-optimized sequence. These errors have been corrected in the HTML and PDF versions of the article, and the correct sequence is provided.