Improved deconvolution of very weak confocal signals.
Improved deconvolution of very weak confocal signals.
复制标题
DOI：
10.12688/f1000research.11773.2
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Glick BS
中科院分区：
文献类型：
--
作者：
Day KJ;La Rivière PJ;Chandler T;Bindokas VP;Ferrier NJ;Glick BS
反卷积通常用于锐化荧光图像，但当信噪比较低时，主要好处是降低噪声并使荧光结构的外观更平滑。 3D 延时 (4D) 共焦图像集可以通过反卷积来改进。然而，当共焦信号非常弱时，流行的惠更斯反卷积软件会擦除原始数据中清晰可见的荧光结构。我们发现这个问题可以通过用高斯模糊对光学部分进行预过滤来避免。对真实数据和模拟数据的分析表明，高斯模糊预滤波器保留了有意义的信号，同时能够去除背景噪声。这种方法非常简单，并且允许惠更斯在 4D 成像条件下使用，从而最大限度地减少光损伤。
Deconvolution is typically used to sharpen fluorescence images, but when the signal-to-noise ratio is low, the primary benefit is reduced noise and a smoother appearance of the fluorescent structures. 3D time-lapse (4D) confocal image sets can be improved by deconvolution. However, when the confocal signals are very weak, the popular Huygens deconvolution software erases fluorescent structures that are clearly visible in the raw data. We find that this problem can be avoided by prefiltering the optical sections with a Gaussian blur. Analysis of real and simulated data indicates that the Gaussian blur prefilter preserves meaningful signals while enabling removal of background noise. This approach is very simple, and it allows Huygens to be used with 4D imaging conditions that minimize photodamage.