单个量子点的光学性质研究及其在超高分辨率定位上的应用
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摘要
量子点(Quantum Dots, QDs)是一种具有诸多优良光学特性的纳米荧光探针,已在化学分析、生物传感、分子影像等领域得到了广泛应用。单个量子点的光学性质研究有望发现一些宏观方法不能发现的实验现象,为改善其光学性能提供思路,也有助于更好的将其应用于各领域。本论文以宽场荧光显微成像技术为主要研究手段,以单个量子点为研究对象,发现了非激发光漂白量子点的行为,提出了影响量子点氧化过程的因素,并将量子点应用于单分子计数和超高分辨率定位,具体内容如下：
一、量子点在非激发光照明下的光漂白行为研究
本章以核壳量子点(Core/shell, CdSe/ZnS)为研究对象,用非激发光连续照射量子点一段时间,结果发现在单分子和系综两个层面上量子点均能被非激发光漂白。这说明即使没有提供足够高的能量使量子点价带的电子跃迁到导带,量子点的光漂白反应依然发生。非激发光下的光漂白动力学与激发光下光漂白动力学相同,均为连续一级基元反应。
二、单个核壳量子点的氧化动力学研究
量子点光谱蓝移是量子点氧化后核尺寸减小的结果。本章利用光谱成像的方法,监测了量子点光谱蓝移的过程,也就相当于监测了量子点的氧化过程。在考察了激发光波长与光强度对蓝移过程的影响后,我们认为量子点的蓝移幅度取决于量子点的激发光波长和量子点的初始尺寸,而与激发光强度无关。量子点的氧化速率正比于激发光的强度,激发光波长越短,量子点的氧化速率越快。
三、量子点团聚程度的测量
纳米颗粒团聚程度的实时测量在技术层面上是一个难点。本章提出了一种基于量子点光谱蓝移和光漂白不同步性的测量量子点团聚体中量子点个数的方法。通过观测量子点团聚物一级条纹的分裂过程,计量出团聚体中量子点的个数。
四、多色和同色量子点复合物的超高分辨率定位技术
多色量子点的超高分辨定位是根据不同颜色量子点的光谱位置不同而识别闪烁的量子点。通过监测双色或多色量子点复合物中个体量子点一级光谱的闪烁,分别定位两种颜色的QD,从而测量出双色量子点间小于衍射极限的距离。
同色量子点的超高分辨率定位是基于量子点光谱蓝移的不同步和量子点漂白的不同步,识别闪烁的量子点,并分别定位。
As novel fluorescent nanomaterials, quantum dots have played important roles in many fileds, such as chemical analysis, biology sensor, molecular imaging, owing to their excellent optical properties. Studies on the properties of single quantum dot can help to find new experimental phenomenology, which can't be found in the ensemble-approach; provide a mentality to improve the properties of quantum dots; contribute to a better application of QDs in various fields. Utilizing the wide-field fluorescence microscopy as the main study approach and taking single QD as research object, this dissertation showed photobleaching of QDs by non-excitation light, presented the influence factor of oxidation process, and applied the QDs to single molecule counting and super-resolution localization. The main work were as follows:
1. Photobleaching of Quantum Dots by Non-excitation Light
Core/shell quantum dots are found to be photobleached by non-excitation wavelength light at the single molecule level and ensemble level. That means even the provided photons have not enough energy to pump electrons into conductive band, the photoreaction of QD for photobleaching still occurs but with slower rate comparing with photobleaching by excitation light. The photodynamic of photobleaching under the non-excitation illumination also follows the consecutive elementary reactions as the behavior of excitation wavelength irradiation.
2. Oxidation Dynamic of Single CdSe/ZnS Quantum Dots
QDs photo-spectal blue shift are caused by an overall size reduction of the QDs due to photooxidation. In this chapter, we utilized the spectral imaging to monitor the process of bule shift, that equals to monitor the QDs oxidation process. After investigated the influence of excitation wavelengh and intensity on Core/shell CdSe/ZnS qauntum dots photo-spectral blue shift, we found that the extent of the blue shift strongly depends on the excitation wavelength and QDs initial size, but not depends on the excitaion intensity. The core oxidation rate is almost proportional to the excitation intensity, shorter excitation wavelength induces faster oxidation rate.
3. A meathod for measuring extent of QDs aggregation
Real-time measurement of the extent of nanopaticle aggregation is a difficult technical problem. In this chapter, we described a simple and accurate stoichiometry method to count the number of QDs among aggregated QDs by recording the process of its spectral buling and photobleaching. Observing aggregated QDs first-order spectral split, the number of spectral split displays the number of QDs.
4. Superlocalization Technique of Multi-color and same-color Quantum Dot Complex
Superlocalization technique of multicolor quantum dot complex is baesd on different spectral position at different color QDs to monitor the QDs blinking. Tracing the first order spectral of multicolor and dualcolor QDs complex, we realized different color QDs localization, separately, and measured the distance between the two QDs, that is less than diffraction limit distance.
Baesd on the QDs spectral bule shift and photobleaching asynchronism, a simple superlocalization technique of same-color QDs complex were developed by identifying QDs blinking.
一、量子点在非激发光照明下的光漂白行为研究
本章以核壳量子点(Core/shell, CdSe/ZnS)为研究对象,用非激发光连续照射量子点一段时间,结果发现在单分子和系综两个层面上量子点均能被非激发光漂白。这说明即使没有提供足够高的能量使量子点价带的电子跃迁到导带,量子点的光漂白反应依然发生。非激发光下的光漂白动力学与激发光下光漂白动力学相同,均为连续一级基元反应。
二、单个核壳量子点的氧化动力学研究
量子点光谱蓝移是量子点氧化后核尺寸减小的结果。本章利用光谱成像的方法,监测了量子点光谱蓝移的过程,也就相当于监测了量子点的氧化过程。在考察了激发光波长与光强度对蓝移过程的影响后,我们认为量子点的蓝移幅度取决于量子点的激发光波长和量子点的初始尺寸,而与激发光强度无关。量子点的氧化速率正比于激发光的强度,激发光波长越短,量子点的氧化速率越快。
三、量子点团聚程度的测量
纳米颗粒团聚程度的实时测量在技术层面上是一个难点。本章提出了一种基于量子点光谱蓝移和光漂白不同步性的测量量子点团聚体中量子点个数的方法。通过观测量子点团聚物一级条纹的分裂过程,计量出团聚体中量子点的个数。
四、多色和同色量子点复合物的超高分辨率定位技术
多色量子点的超高分辨定位是根据不同颜色量子点的光谱位置不同而识别闪烁的量子点。通过监测双色或多色量子点复合物中个体量子点一级光谱的闪烁,分别定位两种颜色的QD,从而测量出双色量子点间小于衍射极限的距离。
同色量子点的超高分辨率定位是基于量子点光谱蓝移的不同步和量子点漂白的不同步,识别闪烁的量子点,并分别定位。
As novel fluorescent nanomaterials, quantum dots have played important roles in many fileds, such as chemical analysis, biology sensor, molecular imaging, owing to their excellent optical properties. Studies on the properties of single quantum dot can help to find new experimental phenomenology, which can't be found in the ensemble-approach; provide a mentality to improve the properties of quantum dots; contribute to a better application of QDs in various fields. Utilizing the wide-field fluorescence microscopy as the main study approach and taking single QD as research object, this dissertation showed photobleaching of QDs by non-excitation light, presented the influence factor of oxidation process, and applied the QDs to single molecule counting and super-resolution localization. The main work were as follows:
1. Photobleaching of Quantum Dots by Non-excitation Light
Core/shell quantum dots are found to be photobleached by non-excitation wavelength light at the single molecule level and ensemble level. That means even the provided photons have not enough energy to pump electrons into conductive band, the photoreaction of QD for photobleaching still occurs but with slower rate comparing with photobleaching by excitation light. The photodynamic of photobleaching under the non-excitation illumination also follows the consecutive elementary reactions as the behavior of excitation wavelength irradiation.
2. Oxidation Dynamic of Single CdSe/ZnS Quantum Dots
QDs photo-spectal blue shift are caused by an overall size reduction of the QDs due to photooxidation. In this chapter, we utilized the spectral imaging to monitor the process of bule shift, that equals to monitor the QDs oxidation process. After investigated the influence of excitation wavelengh and intensity on Core/shell CdSe/ZnS qauntum dots photo-spectral blue shift, we found that the extent of the blue shift strongly depends on the excitation wavelength and QDs initial size, but not depends on the excitaion intensity. The core oxidation rate is almost proportional to the excitation intensity, shorter excitation wavelength induces faster oxidation rate.
3. A meathod for measuring extent of QDs aggregation
Real-time measurement of the extent of nanopaticle aggregation is a difficult technical problem. In this chapter, we described a simple and accurate stoichiometry method to count the number of QDs among aggregated QDs by recording the process of its spectral buling and photobleaching. Observing aggregated QDs first-order spectral split, the number of spectral split displays the number of QDs.
4. Superlocalization Technique of Multi-color and same-color Quantum Dot Complex
Superlocalization technique of multicolor quantum dot complex is baesd on different spectral position at different color QDs to monitor the QDs blinking. Tracing the first order spectral of multicolor and dualcolor QDs complex, we realized different color QDs localization, separately, and measured the distance between the two QDs, that is less than diffraction limit distance.
Baesd on the QDs spectral bule shift and photobleaching asynchronism, a simple superlocalization technique of same-color QDs complex were developed by identifying QDs blinking.
引文
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