Quantum approach for nanoparticle fluorescence by sub‐ns photon detection
Quantum approach for nanoparticle fluorescence by sub‐ns photon detection
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通过亚纳光子检测实现纳米粒子荧光的量子方法
DOI：
10.1002/cyto.a.24310
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Robinson, J. Paul
中科院分区：
文献类型：
--
作者：
Yamamoto, Masanobu;Robinson, J. Paul
明确的检测和分析纳米颗粒大小的样品，如细胞外囊泡或病毒，可能对潜在的疾病诊断很重要。然而，使用传统的流式细胞术光学方法来评估这种小颗粒是相当具有挑战性的。原因是粒子小于衍射极限，使得检测困难。另一种方法是通过附着在纳米颗粒上的共轭荧光染料进行荧光检测；在这种情况下的挑战是对隐藏在高背景光子计数中的极少量发射光子的检测所施加的限制。发射的荧光由众所周知的方程kf = σa I Q描述，该方程将发射的荧光率（kf）（光子/s）描述为分子吸收截面（σa），激发强度(I)和量子产率(Q)的乘积。此外，激发率等于1/t，这是代表流式细胞术中最典型的共轭荧光分子的几个ns寿命的倒数。我们最近开发了一种亚ns光子传感器，它比大多数荧光寿命都快，因为亚ns速度是分离单个发射光子的一个至关重要的参数。根据我们对典型商用流式细胞仪的荧光和背景水平的观察，很明显，背景的一个重要组成部分是由水分子振动引起的。因此，了解构成流式细胞术测量信号的所有成分将有助于定义我们目前所说的“背景信号”。我们试图定义一个理论模型来试图解开这些问题。这个模型是基于在没有水分子的情况下使用反射干燥表面。我们的目标是确定是否有可能最小化背景和增强信号，并提供有关收集到的信号的贡献成分的有价值的信息。为了测试这个模型，我们在最小背景的反射表面上测试了一个直径为50纳米的干燥颗粒。虽然这显然不是一个标准的生物系统，但我们的结果表明，这种量子方法密切遵循已建立的光子基理论。我们的目标是定义实用纳米粒子荧光分析的参数，同时增强我们对背景特性贡献的认识。
Well defined detection and analysis of nanoparticle‐sized samples such as extracellular vesicles or viruses may be important for potential disease diagnostics. However, using conventional flow‐cytometry optical methods to evaluate such small particles is quite challenging. The reason is that the particle is smaller than the diffraction limit, making detection difficult. An alternative approach is fluorescence detection via conjugated fluorochromes attached to the nanoparticles; the challenge in this case is the limitation imposed upon detection of a very small number of emitted photons buried in high background photon counts. Emitted fluorescence is described by the well‐known equation kf = σa I Q, which describes the emitted fluorescence rate (kf) (photons/s) as the multiplication of molecular absorption cross section(σa), excitation intensity (I), and quantum yield (Q). In addition, the excitation rate is equal to 1/t, which is the inverse of the lifetime of several ns representing the most typical conjugated fluorescent molecules used in flow cytometry. We recently developed a sub‐ns photon sensor that is faster than most fluorescence lifetimes, since sub‐ns speed is a critically important parameter for the separation of individual emitted photons. Based on our observation of fluorescence and background levels on typical commercial flow cytometers it is evident that a significant component of the background is induced by water‐molecular vibrations. Therefore, understanding what constitutes all the components that contribute to the signals we measure in flow cytometry would help in defining what we currently call “background signals.” We attempted to define a theoretical model to try to unravel these issues. This model was based on use of a reflective dry surface in the absence of water molecules. Our objective was to determine if it is possible to minimize background and enhance signal, and to provide valuable information on the contributing components of the signals collected. In order to test this model, we tested a single dried particle 50 nm in diameter on a reflective surface with minimum background. While this is clearly not a standard biological system, our results suggest that this quantum approach closely follows established photon base theory. Our goal was to define the parameters for practical nanoparticle‐fluorescence analysis while enhancing our knowledge of the contribution of background properties.
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DOI：
--
发表时间：
2017
期刊：
影响因子：
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作者：
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通讯作者：
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DOI：
--
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2008
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DOI：
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发表时间：
2018
期刊：
BiOS
影响因子：
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作者：
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通讯作者：
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DOI：
--
发表时间：
2006
期刊：
影响因子：
--
作者：
R. Tsien;L. Ernst;A. Waggoner
通讯作者：
A. Waggoner
影响因子：
2.9
作者：
WHITE, JC;STRYER, L
通讯作者：
STRYER, L