Quantitative assessment of fluorescent proteins.

Area of Science:

• Biochemistry
• Molecular Biology
• Microscopy

Background:

• Fluorescent proteins (FPs) are essential tools for genetic labeling in cells and molecules, revolutionizing fluorescence microscopy.
• Genetic engineering has yielded a wide spectrum of bright and stable FPs, from blue to red.
• Autofluorescent FPs share a common rigid beta-barrel structure crucial for fluorescence, yet possess unique properties.

Purpose of the Study:

• To quantitatively characterize over 40 popular and high-performing fluorescent proteins.
• To provide a direct comparison of FP properties including brightness, photostability, pH stability, and monomeric characteristics.
• To guide researchers in selecting the optimal FP for specific experimental needs.

Main Methods:

• Quantitative characterization of fluorescent protein properties.
• Focus on brightness, photostability, pH stability, and monomeric status.
• Inclusion of popular and top-performing FPs across the blue to red spectral regions.

Main Results:

• Detailed comparative data on the performance of more than 40 fluorescent proteins.
• Identification of unique advantages and disadvantages for each FP.
• Enabling straightforward comparison to aid in FP selection.

Conclusions:

• No single fluorescent protein is universally optimal; selection depends on the application.
• This quantitative characterization provides a valuable resource for researchers.
• Informed FP choice enhances the success of fluorescence microscopy experiments.