Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 124, Issue 10, Pages 1892-1897Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpcb.9b11028
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Funding
- Ministry of Science and Technology of China [2016YFA0500700]
- National Natural Science Foundation of China [11925406, 21573214, 118723585]
- Collaborative Innovation Program of Hefei Science Center, CAS [2019HSC-CIP004]
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Photoactivatable fluorescent proteins (PA-FPs) are widely used in photoactivated localization microscopy (PALM), and information about their blinking properties is important for precise assembling of PALM images and for quantitative molecular counting with PALM. As the photophysical properties of fluorescent proteins depend sensitively on the local physiochemical environment, we developed a series of procedures to characterize the properties of the PA-FPs in fixed cells. As an example, we characterized the properties of Dendra2 in Escherichia coli and found that they are appreciably different from those in vitro. This highlights the importance of characterizing the photophysical properties of PA-FPs under the same experimental conditions as subsequent PALM imaging or molecular counting experiments. Other PA-FPs could be characterized using this strategy.
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