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Related Experiment Videos

A guide to choosing fluorescent proteins.

Nathan C Shaner1, Paul A Steinbach, Roger Y Tsien

  • 1Department of Pharmacology, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.

Nature Methods
|November 22, 2005
PubMed
Summary
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A wide array of fluorescent proteins (FPs) are now available for biological imaging. This guide offers a unified characterization to help researchers select the best FPs for their specific experimental needs, reducing trial-and-error.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Microscopy

Background:

  • The rapid proliferation of fluorescent proteins (FPs) offers advanced capabilities for biological imaging.
  • Lack of standardized assessment methods complicates FP selection for researchers.
  • Key questions arise regarding optimal FP choice for general use, brightness, and experimental suitability.

Purpose of the Study:

  • To provide a unified characterization of commonly used fluorescent proteins.
  • To guide researchers in selecting appropriate FPs for diverse biological imaging applications.
  • To establish a framework for evaluating FP performance beyond basic brightness.

Main Methods:

  • Comparative analysis of spectral properties (excitation/emission maxima, extinction coefficients, quantum yields).

Related Experiment Videos

  • Assessment of photostability and maturation times under various imaging conditions.
  • Evaluation of FP performance in common cellular contexts and imaging modalities.

    • Main Results:

    • Detailed comparison of brightness, photostability, and spectral profiles for a curated list of FPs.
    • Identification of FPs excelling in specific applications (e.g., high-resolution microscopy, long-term imaging).
    • Quantification of maturation kinetics and potential for aggregation.

    Conclusions:

    • A standardized characterization is crucial for efficient FP selection in biological imaging.
    • Specific FPs demonstrate superior performance for particular experimental requirements.
    • This resource aids researchers in navigating FP diversity and optimizing imaging experiments.