研究目的
研究在超分辨率成像中使用特种探针以突破衍射极限,实现超过200纳米的细胞细节观察。
研究成果
该论文得出结论:随着探针和成像技术的进步,活细胞超分辨率成像有望成为常规技术。研究强调了开发光开关和光转换荧光蛋白(FPs)对多色实验及深度成像技术的重要性。
研究不足
该论文提到了活细胞成像中光毒性的挑战、红色探针相较于绿色探针性能的局限,以及需要推进探针和成像模式的发展以减少光漂白、提高速度并增加从活细胞记录的图像数量。
该论文探讨了多种超分辨率成像技术,如RESOLFT、PALM/STORM、STED和SIM,以及能够从暗态切换至亮态、从一种颜色转换至另一种颜色的特种探针、染料和荧光蛋白(FPs)。研究强调了将探针与成像模式相匹配的重要性,以充分发挥两者的优势,从而实现超越衍射极限的超分辨率成像。
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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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