研究目的
Investigating the use of specialty probes in super-resolution imaging to overcome the diffraction limit and achieve cellular detail beyond 200 nanometers.
研究成果
The paper concludes that super-resolution imaging with live cells has the potential to become routine with advances in probes and imaging modalities. It emphasizes the importance of developing photoswitchable and photoconvertible FPs for multicolor experiments and deep imaging techniques.
研究不足
The paper mentions the challenges of phototoxicity in live-cell imaging, the limited performance of red probes compared to green ones, and the need for advances in probes and imaging modalities to reduce photobleaching and increase speed and the number of images recorded from living cells.
The paper discusses various super-resolution imaging techniques such as RESOLFT, PALM/STORM, STED, and SIM, and the use of specialty probes, dyes, and fluorescent proteins (FPs) that can switch from dark to light and from one color to another. It highlights the importance of matching probes to imaging modalities to maximize the strengths of both for achieving super-resolution imaging beyond the diffraction limit.
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rsEGFP2
Used in RESOLFT microscopy for its high stability against bleaching.
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Dronpa
A green fluorescent protein used in super-res imaging, capable of switching between fluorescent and non-fluorescent states.
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rsFusionRed
A red photoswitchable fluorescent protein used in MoNaLISA and RESOLFT for live-cell imaging.
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mEos3.2
Preferred for PALM/STORM due to its photoconvertible properties.
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Skylan-S
Suitable for super-resolution optical fluctuation imaging (SOFI).
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Skylan-NS
Pairs well with PA NL-SIM for live-cell super-res imaging.
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Padron
Developed as a partner for Dronpa, used in reversible switching experiments.
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Dreiklang
A YFP-based FP with decoupled on/off switch from fluorescence readout, used in live mammalian cells.
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ORCA-Fusion
Hamamatsu
A camera designed for low-light super-resolution imaging, with low read noise for single-molecule measurements.
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