Weigel Arwed, Schild Detlev, Zeug André
University of Gottingen, Department of Neurophysiology and Cellular Biophysics, Humboldtallee 23, 37073 Gottingen, Germany.
J Biomed Opt. 2009 Jan-Feb;14(1):014022. doi: 10.1117/1.3083439.
The essential feature of the confocal laser scanning microscope (cLSM) is the generation of optical sections by the removal of out-of-focus light. About ten years ago, structured illumination microscopy (SIM) was introduced as an alternative method for obtaining optical sections from biological specimens. Here we compare the resolution of the ApoTome (commercial SIM by Zeiss) to that achieved by a cLSM (Zeiss LSM 510). If fluorescent beads are used as test objects, then the ApoTome will achieve a lower axial resolution than the cLSM. In contrast to that, its lateral resolution scores slightly better. If subresolution homogeneous fluorescent layers are used as test objects, then the ApoTome will achieve a higher axial resolution than the cLSM. The ApoTome's axial resolution is homogeneous over the field-of-view while that of the cLSM changes markedly. Finally, the anisotropy of the ApoTome's resolution was found to be negligible for standard applications while its capability to resolve fine structures within stained tissue slices is limited to one or two cell layers and thus worse than in the cLSM.
共聚焦激光扫描显微镜(cLSM)的基本特征是通过去除离焦光来生成光学切片。大约十年前,结构照明显微镜(SIM)作为从生物标本获取光学切片的一种替代方法被引入。在此,我们将ApoTome(蔡司公司的商用SIM)的分辨率与cLSM(蔡司LSM 510)所达到的分辨率进行比较。如果使用荧光微球作为测试对象,那么ApoTome的轴向分辨率将低于cLSM。与此相反,其横向分辨率略好一些。如果使用亚分辨率均匀荧光层作为测试对象,那么ApoTome的轴向分辨率将高于cLSM。ApoTome的轴向分辨率在整个视野范围内是均匀的,而cLSM的轴向分辨率则有显著变化。最后,发现对于标准应用而言,ApoTome分辨率的各向异性可忽略不计,而其分辨染色组织切片内精细结构的能力仅限于一到两个细胞层,因此比cLSM要差。