NEAR-FIELD SCANNING OPTICAL MICROSCOPY (NSOM) - DEVELOPMENT AND BIOPHYSICAL APPLICATIONS
NEAR-FIELD SCANNING OPTICAL MICROSCOPY (NSOM) - DEVELOPMENT AND BIOPHYSICAL APPLICATIONS
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DOI：
10.1016/s0006-3495(86)83640-2
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发表时间：
1986-01-01
影响因子：
3.4
通讯作者：
KRATSCHMER, E
中科院分区：
文献类型：
--
作者：
BETZIG, E;LEWIS, A;KRATSCHMER, E
一种用于高分辨率成像的新方法——近场扫描光学显微镜（NSOM）已经被开发出来。讨论了控制该方法的概念，并描述了构建工作 NSOM 仪器时遇到的技术挑战。提出了两种不同的方法来制造特征良好、高度可重复的亚波长孔径。提供样本一维扫描并将其与测试图案的扫描电子显微照片进行比较。通过比较，确定了 > 1,500Å（即 ≃λ/3.6）的分辨率，这代表着我们朝着 500Å 分辨率的最终目标迈出了重要一步。通过小于 600Å 的孔径观察到荧光，信噪比计算表明荧光成像应该是可行的。然后结合具体的生物学问题讨论这种成像的应用。 NSOM 方法采用非电离可见辐射，可在空气或水环境中用于功能生物系统的无损可视化，其分辨率与扫描电子显微镜相当。
A new method for high-resolution imaging, near-field scanning optical microscopy (NSOM), has been developed. The concepts governing this method are discussed, and the technical challenges encountered in constructing a working NSOM instrument are described. Two distinct methods are presented for the fabrication of well-characterized, highly reproducible, subwavelength apertures. A sample one-dimensional scan is provided and compared to the scanning electron micrograph of a test pattern. From this comparison, a resolution of > 1,500Å (i.e., ≃λ/3.6) is determined, which represents a significant step towards our eventual goal of 500Å resolution. Fluorescence has been observed through apertures smaller than 600Å and signal-to-noise calculations show that fluorescent imaging should be feasible. The application of such imaging is then discussed in reference to specific biological problems. The NSOM method employs nonionizing visible radiation and can be used in air or aqueous environments for nondestructive visualization of functioning biological systems with a resolution comparable to that of scanning electron microscopy.