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相衬显微镜上的分子对比。

Molecular contrast on phase-contrast microscope.

机构信息

Department of Physics, The University of Tokyo, Tokyo, 113-0033, Japan.

Department of Neurology, The University of Tokyo, Tokyo, 113-0033, Japan.

出版信息

Sci Rep. 2019 Jul 18;9(1):9957. doi: 10.1038/s41598-019-46383-6.

Abstract

An optical microscope enables image-based findings and diagnosis on microscopic targets, which is indispensable in many scientific, industrial and medical settings. A standard benchtop microscope platform, equipped with e.g., bright-field and phase-contrast modes, is of importance and convenience for various users because the wide-field and label-free properties allow for morphological imaging without the need for specific sample preparation. However, these microscopes never have capability of acquiring molecular contrast in a label-free manner. Here, we develop a simple add-on optical unit, comprising of an amplitude-modulated mid-infrared semiconductor laser, that is attached to a standard microscope platform to deliver the additional molecular contrast of the specimen on top of its conventional microscopic image, based on the principle of photothermal effect. We attach this unit, termed molecular-contrast unit, to a standard phase-contrast microscope, and demonstrate high-speed label-free molecular-contrast phase-contrast imaging of silica-polystyrene microbeads mixture and molecular-vibrational spectroscopic imaging of HeLa cells. Our simple molecular-contrast unit can empower existing standard microscopes and deliver a convenient accessibility to the molecular world.

摘要

光学显微镜能够基于微观目标进行基于图像的发现和诊断,在许多科学、工业和医疗环境中都不可或缺。配备明场和相差模式等功能的标准台式显微镜平台对于各种用户来说都非常重要和方便,因为其宽场和无标记的特性允许对样品进行无需特定制备的形态成像。然而,这些显微镜从未具有以无标记方式获取分子对比度的能力。在这里,我们开发了一个简单的附加光学单元,包括一个振幅调制的中红外半导体激光器,该单元连接到标准显微镜平台上,基于光热效应的原理,在常规显微镜图像的基础上为标本提供额外的分子对比度。我们将这个单元称为分子对比度单元,附加到标准相差显微镜上,并演示了硅聚苯乙烯微球混合物的高速无标记分子对比度相差成像以及 HeLa 细胞的分子振动光谱成像。我们的简单分子对比度单元可以为现有标准显微镜提供增强功能,并为进入分子世界提供便捷的途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9700/6637114/b978c5964bd4/41598_2019_46383_Fig1_HTML.jpg

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