Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications - I. Theory
Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications - I. Theory
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DOI：
10.1006/abio.1998.2759
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发表时间：
1998-09-10
影响因子：
2.9
通讯作者：
Yguerabide, EE
中科院分区：
文献类型：
--
作者：
Yguerabide, J;Yguerabide, EE
当用白色光照射时，亚微观金颗粒悬浮液散射彩色光，并且我们已经观察到光散射金颗粒悬浮液具有与荧光溶液相同的外观。因此，当用窄的白色光束照射时，40 nm的金溶胶显示出透明（不浑浊）的绿色散射光（廷德尔）光束，并具有与发荧光的荧光素溶液相同的外观。这些，以及其他，观察向我们表明，在一般情况下，光散射颗粒可以被视为荧光类似物，并作为荧光类似物示踪剂在免疫和DNA探针测定以及使用。如细胞和分子生物学研究。光散射颗粒在这些应用中是有利的，因为诸如金和银的颗粒具有非常高的光散射能力，这使得这些颗粒在低至10（-16）M的颗粒浓度下通过光散射容易地检测。散射光可以通过肉眼检测到，用于定性测量，或者用简单的光敏检测器检测到，用于定量测量。此外，单个颗粒可以通过眼睛或使用具有适当照明系统的简单光学显微镜的摄像机容易地检测到。此外，可以容易地合成散射蓝色、绿色、黄色、橙子或红色光的亚微观颗粒。抗体、DNA探针和其他示踪物质可以很容易地附着在金和其他粒子上，而不会改变它们的光散射特性。在这篇文章中，我们提出的理论，它允许一个预测的光散射特性的颗粒的不同尺寸和组成，并确定那些颗粒的尺寸和组成，似乎最适合于特定的应用。此外，我们计算了不同尺寸和组成的颗粒的摩尔消光系数和发射效率，这使我们能够将这些颗粒的发光能力与众所周知的荧光示踪剂的发光能力进行比较。例如，60 nm的金颗粒相当于约3 × 10（5）个荧光素分子。使用光散射颗粒作为荧光类似物示踪剂，可以开发非常简单、易于使用、低成本、超灵敏的免疫和DNA探针测定。如文章所述，使用光学显微镜技术，在适当的照明下，可以在细胞表面和细胞内部检测到单个颗粒。在高颗粒密度下，颗粒标记的细胞具有与荧光细胞相同的外观。(C)北京：科学出版社.
Submicroscopic gold particle suspensions scatter colored light when illuminated with white light, and we have observed that a light-scattering gold particle suspension has the same appearance as a fluorescing solution. Thus, when illuminated by a narrow beam of white light, a 40-nm gold sol displays a clear (not cloudy), green scattered light (Tyndall) beam and has the same appearance as a fluorescing fluorescein solution. These, as well as other, observations have suggested to us that, in general, light-scattering particles can be treated as fluorescent analogs and used as fluorescent analog tracers in immuno- and DNA probe assays as well. as in cell, and molecular biology studies. Light-scattering particles are advantageous in these applications because particles such as gold and silver have very high light-scattering powers, which allows these particles to be easily detected, by light-scattering, at particle concentrations as low as 10(-16) M. The scattered light can be detected by the unaided eye for qualitative measurements or with a simple light-sensitive detector for quantitative measurements. Moreover, individual particles can be easily detected by eye or a video camera using a simple light microscope with a proper illuminating system. In addition, submicroscopic particles which scatter blue, green, yellow, orange, or red light can be readily synthesized. Antibodies, DNA probes, and other tracer substances can be readily attached to gold and other particles without altering their light-scattering properties. In this article we present the theory which allows one to predict the light-scattering properties of particles of different sizes and compositions and identify those particle sizes and compositions which appear most adequate for particular applications. Furthermore, we calculate molar extinction coefficients and emission efficiencies for particles of different sizes and compositions which allows us to compare the light-producing powers of these particles with those of well known fluorescent tracers. A 60-nm gold particle, for example, is equivalent to about 3 x 10(5) fluorescein molecules. Very simple, easy to use, low-cost, ultrasensitive immuno- and DNA probe assays can be developed using light-scattering particles as fluorescent analog tracers. Single particles can be detected on cell surfaces and inside cells using light microscopy techniques with proper illumination as described in the article. At high particle densities, particle-labeled cells have the same appearance as fluorescent cells. (C) 1998 Academic Press.