Exploring the Potential of Airyscan Microscopy for Live Cell Imaging
Exploring the Potential of Airyscan Microscopy for Live Cell Imaging
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DOI：
10.3390/photonics4030041
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发表时间：
2017-09-01
期刊：
影响因子：
2.4
通讯作者：
Fritzsche, Marco
中科院分区：
文献类型：
--
作者：
Korobchevskaya, Kseniya;Lagerholm, B. Christoffer;Fritzsche, Marco
生物研究越来越需要使用非侵入性和超灵敏成像技术。Airyscan技术是最近开发的，用于弥合传统共焦显微镜和超分辨率显微镜之间的差距。该技术结合了共焦成像与0.2艾里单位针孔，去卷积和像素重新分配原则，以提高空间分辨率和信噪比，而不增加激发功率和采集时间。在这里，我们提出了一个详细的研究评估性能的Airyscan相比，共聚焦显微镜成像的各种参考样品和生物标本与不同的采集和处理参数。我们发现，处理后的Airyscan图像在默认的反卷积设置具有类似于传统的共焦成像的空间分辨率与针孔设置为0.2艾里单位，但具有显着改善的信号噪声比。通过使用增强的去卷积滤波器设置可以实现空间分辨率的进一步增益，但信噪比会稳定损失，在更极端的设置下会导致显著的数据损失和图像失真。
Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment principle in order to enhance both the spatial resolution and signal-to-noise-ratio without increasing the excitation power and acquisition time. Here, we present a detailed study evaluating the performance of Airyscan as compared to confocal microscopy by imaging a variety of reference samples and biological specimens with different acquisition and processing parameters. We found that the processed Airyscan images at default deconvolution settings have a spatial resolution similar to that of conventional confocal imaging with a pinhole setting of 0.2 Airy Units, but with a significantly improved signal-to-noise-ratio. Further gains in the spatial resolution could be achieved by the use of enhanced deconvolution filter settings, but at a steady loss in the signal-to-noise ratio, which at more extreme settings resulted in significant data loss and image distortion.