Castello Marco, Sheppard Colin J R, Diaspro Alberto, Vicidomini Giuseppe
Opt Lett. 2015 Nov 15;40(22):5355-8. doi: 10.1364/OL.40.005355.
Confocal scanning microscopy (CSM) is the most widely used modern optical microscopy technique. Theoretically, it allows the diffraction barrier to be surpassed by a factor of 2, but practically this improvement is sacrificed to obtain a good signal-to-noise ratio (SNR). Image scanning microscopy (ISM) solves this limitation but, in the current implementations, the system complexity is increased and the versatility of CSM is reduced. Here we show that ISM can be straightforwardly implemented by substituting the single point detector of a confocal microscope with a quadrant detector of the same size, thus using a small number of detector elements. This implementation offers resolution close to the CSM theoretical value and improves the SNR by a factor of 1.5 with respect to the CSM counterpart without losing the optical sectioning capability and the system versatility.
共聚焦扫描显微镜(CSM)是应用最为广泛的现代光学显微镜技术。理论上,它能将衍射极限突破两倍,但实际上为了获得良好的信噪比(SNR),这种改进会被牺牲。图像扫描显微镜(ISM)解决了这一局限性,但在当前的实现方式中,系统复杂性增加,CSM的通用性降低。在此我们表明,通过用相同尺寸的象限探测器替代共聚焦显微镜的单点探测器,从而使用少量探测器元件,就能直接实现ISM。这种实现方式提供了接近CSM理论值的分辨率,并且相对于CSM的同类设备,在不损失光学切片能力和系统通用性的情况下,将信噪比提高了1.5倍。