Reduced Dyes Enhance Single-Molecule Localization Density for Live Superresolution Imaging
Reduced Dyes Enhance Single-Molecule Localization Density for Live Superresolution Imaging
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DOI：
10.1002/cphc.201301004
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发表时间：
2014-03-17
期刊：
影响因子：
2.9
通讯作者：
Manley, Suliana
中科院分区：
文献类型：
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作者：
Carlini, Lina;Benke, Alexander;Manley, Suliana
细胞可渗透的罗丹明染料被 NaBH4 还原猝灭成非荧光无色罗丹明形式。淬灭是可逆的，当染料被氧化时，它们的荧光会恢复。在活细胞中，氧化会自发发生，与未修饰的染料相比，可导致单分子定位密度高出十倍，并且每个定位产生更多光子。这两个参数直接影响可实现的分辨率，我们看到使用减少的染料拍摄的活细胞点定位超分辨率图像的质量显着提高。这些改进进一步提高了单分子跟踪实验的轨迹密度。
Cell-permeable rhodamine dyes are reductively quenched by NaBH4 into a non-fluorescent leuco-rhodamine form. Quenching is reversible, and their fluorescence is recovered when the dyes are oxidized. In living cells, oxidation occurs spontaneously, and can result in up to ten-fold higher densities of single molecule localizations, and more photons per localization as compared with unmodified dyes. These two parameters directly impact the achievable resolution, and we see a significant improvement in the quality of live-cell point-localization super-resolution images taken with reduced dyes. These improvements carry over to increase the density of trajectories for single-molecule tracking experiments.