Pixel reassignment in image scanning microscopy: a re-evaluation

针孔(光学) 显微镜 光学 显微镜 共焦 4Pi显微镜 材料科学 共焦显微镜 扫描共焦电子显微镜 人工智能 分辨率(逻辑) 点扩散函数 光学显微镜 探测器 像素 计算机科学 图像分辨率 物理 扫描电子显微镜 常规透射电子显微镜 扫描透射电子显微镜
作者
Colin J. R. Sheppard,Marco Castello,Giorgio Tortarolo,Takahiro Deguchi,Sami Koho,Giuseppe Vicidomini,Alberto Diaspro
出处
期刊:Journal of the Optical Society of America [Optica Publishing Group]
卷期号:37 (1): 154-154 被引量:38
标识
DOI:10.1364/josaa.37.000154
摘要

Image scanning microscopy is a technique based on confocal microscopy, in which the confocal pinhole is replaced by a detector array, and the resulting image is reconstructed, usually by the process of pixel reassignment. The detector array collects most of the fluorescent light, so the signal-to-noise ratio is much improved compared with confocal microscopy with a small pinhole, while the resolution is improved compared with conventional (wide-field) microscopy. In previous studies, it has usually been assumed that pixels should be reassigned by a constant factor, to a point midway between the illumination and detection spots. Here it is shown that the peak intensity of the effective point spread function (PSF) can be further increased by 4% by a new choice of the pixel reassignment factor. For an array of two Airy units, the peak of the effective PSF is 1.90 times that of a conventional microscope, and the transverse resolution is 1.53 times better. It is confirmed that image scanning microscopy gives optical sectioning strength identical to that of a confocal microscope with a pinhole equal to the size of the detector array. However, it is shown that image scanning microscopy exhibits axial resolution superior to a confocal microscope with a pinhole the same size as the detector array. For a two-Airy-unit array, the axial resolution is 1.34 times better than in a conventional microscope for the standard reassignment factor, and 1.28 times better for the new reassignment factor. The axial resolution of a confocal microscope with a two-Airy-unit pinhole is only 1.04 times better than conventional microscopy. We also examine the signal-to-noise ratio of a point object in a uniform background (called the detectability), and show that it is 1.6 times higher than in a confocal microscope.
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