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Photon scanning-tunneling microscopy of unstained mammalian cells and chromosomes

机译：未染色的哺乳动物细胞和染色体的光子扫描隧道显微镜

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The photon scanning-tunneling microscope (PSTM) yields optical topographical images of samples that are thin or that are transparent at the wavelength used. A range of sample sizes can be imaged extending to well below the diffraction Limit for sufficiently flat samples. But samples of the order of several to many micrometers in size can be analyzed with less-refined resolution if total internal reflection can be made to occur in the sample. We used the PSTM to examine the optical topography of mouse and human cells and of chromosomes that are unstained. Our objectives were to demonstrate the images as an alternative to conventional microscopy and to provide a sample-preparation methodology that will later permit localized, simultaneous fluorescence or absorption spectroscopy with the signals collected by the probe tip. Furthermore, the PSTM's ability to produce optical profiles in air and in water was tested to establish the basis for future investigation of possible abnormalities in the chromosomes. That is, we considered both physical and biological objectives. To this end we utilized the 442-nm line of a He-Cd laser as well as the 633-nm line from a He-Ne laser, the resulting image quality being tested partly to ascertain the increased effects of scattering at the smaller wavelength. It is shown that adequate resolution and signal-to-noise ratio can be obtained with the shorter wavelength Even in the presence of intensity fluctuations from the laser, thus showing that fluorescence and absorption studies can be expected to be practicable. (C) 1998 Optical Society of America OCIS codes: 170.1530, 180.0180, 180.5810. [References: 23]
机译：光子扫描隧道显微镜（PSTM）产生的样品的光学形貌图在使用的波长下较薄或透明。对于足够平坦的样品，可以将样品尺寸范围扩展到远低于衍射极限的范围。但是，如果可以在样品中发生全内反射，则可以以较不精确的分辨率分析大小为几微米到几微米的样品。我们使用PSTM检查了小鼠和人类细胞以及未染色染色体的光学形貌。我们的目标是展示图像作为常规显微镜的替代方法，并提供一种样品制备方法，该方法随后将利用探针尖端收集的信号进行局部同时荧光或吸收光谱分析。此外，测试了PSTM在空气和水中产生光学图谱的能力，为将来研究染色体可能的异常情况奠定了基础。也就是说，我们同时考虑了物理和生物学目标。为此，我们使用了He-Cd激光器的442 nm线以及He-Ne激光器的633 nm线，对所得图像质量进行了部分测试，以确定在较小波长处散射的增强效果。结果表明，即使在存在来自激光器的强度波动的情况下，也可以在较短的波长下获得足够的分辨率和信噪比，从而表明可以预期进行荧光和吸收研究。 （C）1998年美国光学学会OCIS编码：170.1530、180.0180、180.5810。 [参考：23]

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