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A guide to choosing fluorescent proteins


The recent explosion in the diversity of available fluorescent proteins (FPs)1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16 promises a wide variety of new tools for biological imaging. With no unified standard for assessing these tools, however, a researcher is faced with difficult questions. Which FPs are best for general use? Which are the brightest? What additional factors determine which are best for a given experiment? Although in many cases, a trial-and-error approach may still be necessary in determining the answers to these questions, a unified characterization of the best available FPs provides a useful guide in narrowing down the options.

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Thanks to S. Adams for helpful advice on choosing filter sets. N.C.S. is a Howard Hughes Medical Institute Predoctoral Fellow. This work was additionally supported by US National Institutes of Health (NS27177 and GM72033) and Howard Hughes Medical Institutes.

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Correspondence to Roger Y Tsien.

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Competing interests

Fluorescent proteins described in the paper that originate from the Tsien lab are covered by pending US and international patents. These patents, when issued, will be property of the University of California.

Supplementary information

Supplementary Fig. 1

mCherry and Emerald photobleaching curves.

Supplementary Table 1

FPs properties in detail.

Supplementary Table 2

Mutations of AFPs relative to wild-type GFP.

Supplementary Table 3

Starting points for advanced FP applications.

Supplementary Discussion

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Shaner, N., Steinbach, P. & Tsien, R. A guide to choosing fluorescent proteins. Nat Methods 2, 905–909 (2005).

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