Quantitative comparison of spinning disk geometries for PAINT based super-resolution microscopy.
Quantitative comparison of spinning disk geometries for PAINT based super-resolution microscopy.
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基于PAINT的超分辨率显微镜旋转盘几何形状的定量比较。
DOI：
10.1364/boe.459490
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发表时间：
2022-07-01
影响因子：
3.4
通讯作者：
St Johnston, Daniel
中科院分区：
文献类型：
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作者：
Sirinakis, George;Allgeyer, Edward S.;Cheng, Jinmei;St Johnston, Daniel
使用基于DNA或蛋白质的可交换探针的PAINT方法已成为超分辨率成像的流行方法，并已与旋转盘共聚焦显微镜相结合，用于成像较厚的样品。然而，广泛用于常规生物成像的旋转磁盘并没有针对基于paint的应用进行优化，可能会降低分辨率和成像速度。在这里，我们使用肌动蛋白结合肽Lifeact存在的果蝇卵室来研究四种不同旋转盘几何形状的性能。我们发现具有更高光收集效率的磁盘几何形状在基于paint的超分辨率成像中表现更好，因为光子数量增加，随后检测到更多的闪烁事件。
PAINT methods that use DNA- or protein- based exchangeable probes have become popular for super-resolution imaging and have been combined with spinning disk confocal microscopy for imaging thicker samples. However, the widely available spinning disks used for routine biological imaging are not optimized for PAINT-based applications and may compromise resolution and imaging speed. Here, we use Drosophila egg chambers in the presence of the actin-binding peptide Lifeact to study the performance of four different spinning disk geometries. We find that disk geometries with higher light collection efficiency perform better for PAINT-based super-resolution imaging due to increased photon numbers and, subsequently, detection of more blinking events.