Opt Express. 2022 Apr 11;30(8):13825-13838. doi: 10.1364/OE.455621.
Image scanning microscopy (ISM) overcomes the trade-off between spatial resolution and signal volume in confocal microscopy by rearranging the signal distribution on a two-dimensional detector array to achieve a spatial resolution close to the theoretical limit achievable by infinitesimal pinhole detection without sacrificing the detected signal intensity. In this paper, we improved the spatial resolution of ISM in three dimensions by exploiting saturated excitation (SAX) of fluorescence. We theoretically investigated the imaging properties of ISM, when the fluorescence signals are nonlinearly induced by SAX, and show combined SAX-ISM fluorescence imaging to demonstrate the improvement of the spatial resolution in three dimensions. In addition, we confirmed that the SNR of SAX-ISM imaging of fluorescent beads and biological samples, which is one of the challenges in conventional SAX microscopy, was improved.
图像扫描显微镜（ISM）通过重新排列二维探测器阵列上的信号分布来克服共聚焦显微镜中空间分辨率和信号体积之间的权衡，从而实现接近微小孔检测理论极限的空间分辨率，而不会牺牲检测到的信号强度。在本文中，我们通过荧光的饱和激发（SAX）来提高 ISM 的三维空间分辨率。我们从理论上研究了当荧光信号被 SAX 非线性诱导时 ISM 的成像特性，并展示了组合的 SAX-ISM 荧光成像，以证明三维空间分辨率的提高。此外，我们证实了荧光珠和生物样品的 SAX-ISM 荧光成像的 SNR（信噪比）得到了提高，这是传统 SAX 显微镜面临的挑战之一。
