暗場(chǎng)散射成像在刺激響應(yīng)型反應(yīng)監(jiān)控中的應(yīng)用及高性能成像系統(tǒng)研究
發(fā)布時(shí)間：2018-07-15 14:34
【摘要】：暗場(chǎng)散射成像技術(shù),作為一種高對(duì)比度和非掃描的光學(xué)成像技術(shù),被廣泛的應(yīng)用于分析傳感、生物過(guò)程示蹤以及反應(yīng)監(jiān)控等領(lǐng)域。單納米顆粒等離子體探針,具有傳統(tǒng)散射探針無(wú)法比擬的優(yōu)勢(shì),如穩(wěn)定且強(qiáng)的光學(xué)信號(hào),精準(zhǔn)的局域信息以及易于調(diào)控的性質(zhì),使得散射成像在納米尺度的動(dòng)態(tài)反應(yīng)監(jiān)控和空間分辨方面具有廣闊的應(yīng)用前景。等離子體耦合引起的散射增強(qiáng)和光譜移動(dòng)也被廣泛應(yīng)用于納米標(biāo)尺的開(kāi)發(fā)和生物醫(yī)學(xué)分析檢測(cè)。然而,目前利用暗場(chǎng)散射成像來(lái)監(jiān)控和研究反應(yīng)過(guò)程的報(bào)道為數(shù)不多,對(duì)于刺激響應(yīng)型的可控反應(yīng)的監(jiān)控更是鮮有報(bào)道,可能是由于缺乏適當(dāng)?shù)难芯磕Ｐ鸵约俺上裥盘?hào)和動(dòng)態(tài)反應(yīng)難以建立有效的銜接。目前,暗場(chǎng)成像技術(shù)自身仍存在一定的局限性,包括可見(jiàn)度在一定程度上受限于較低的光源利用率,以及衍射極限的束縛使得的空間分辨受限。本論文圍繞上述的研究難點(diǎn)和問(wèn)題,借鑒前人的工作基礎(chǔ),在散射成像監(jiān)控動(dòng)態(tài)反應(yīng)過(guò)程以及成像性能改進(jìn)方面開(kāi)展了如下具體的工作：1.暗場(chǎng)散射成像對(duì)配合物納米藥物載體的酸敏感性的研究。利用有機(jī)配體1,1’-(1,4-丁基)二咪唑和亞鐵離子制備了配位聚合物微球,同時(shí)實(shí)現(xiàn)了對(duì)抗癌藥物鹽酸阿霉素的高效原位包覆。通過(guò)反應(yīng)溶劑的調(diào)控,可以獲得最優(yōu)的高達(dá)98%的藥物負(fù)載率和近40%的藥物負(fù)載效率。此納米藥物載體由于其自身具有的pH敏感性,以及經(jīng)表面硅烷化后進(jìn)行的癌細(xì)胞靶向劑葉酸的修飾,被用于抗癌藥物的靶向傳輸和pH敏感的緩釋和控釋。熒光成像結(jié)果表明此載體具有靶向作用,SEM對(duì)各時(shí)間點(diǎn)的微球形貌研究結(jié)果可以證明其酸敏感性,卻無(wú)法對(duì)單顆粒水平進(jìn)行連續(xù)實(shí)時(shí)監(jiān)控,而暗場(chǎng)散射成像則實(shí)現(xiàn)了對(duì)酸敏感的微球載體降解過(guò)程的單顆粒水平的實(shí)時(shí)原位監(jiān)控,直觀地考察了藥物載體的酸敏感性。2.單顆粒散射成像對(duì)光致化學(xué)鍵斷裂反應(yīng)的實(shí)時(shí)監(jiān)控。將球形和棒狀兩種形體的銀納米顆粒作為暗場(chǎng)成像下的檢測(cè)探針,實(shí)現(xiàn)了對(duì)共價(jià)連接的銀-二硫代氨基甲酸(Ag-DTC)化學(xué)鍵的光敏感性的實(shí)時(shí)監(jiān)控。通過(guò)觀察Ag-DTC斷裂后所伴隨產(chǎn)生的硫氫根介導(dǎo)形成的Ag@Ag2S核殼結(jié)構(gòu)帶來(lái)的散射信號(hào)紅移,發(fā)現(xiàn)此化學(xué)鍵的斷裂可以有效地被光驅(qū)動(dòng),同時(shí)對(duì)另外幾個(gè)重要的影響因素,比如pH,溶劑極性以及還原劑等,也進(jìn)行了全面的考察。借助軟件進(jìn)行成像數(shù)據(jù)分析,可以精確地發(fā)現(xiàn)球形AgNPs探針的散射強(qiáng)度在反應(yīng)監(jiān)控過(guò)程中,存在一個(gè)先升高后降低的過(guò)程,單顆粒散射光譜也證實(shí)了這一現(xiàn)象。對(duì)于光的驅(qū)動(dòng)機(jī)制解釋為光生熱電子克服Ag-DTC斷鍵的能壘,同時(shí)結(jié)合銀的較高的氧化還原活性,從而表現(xiàn)出Au-DTC所不具有的光敏感性。3.基于表面等離激元“光濃縮”效應(yīng)和光路改裝提升成像可見(jiàn)度的研究。目前,暗場(chǎng)成像主要采用斜射照明,通過(guò)入射光與樣品的散射光的空間分離獲得優(yōu)異的成像對(duì)比度。如此一來(lái),光源的利用效率比較低,從而使得弱散射信號(hào)的可見(jiàn)度受損,同時(shí)對(duì)光源的功率要求較高。等離子體納米顆粒具有大于物理截面的散射截面,從而可以將光集中到顆粒附近區(qū)域,從而散射出很強(qiáng)的光。在干式暗場(chǎng)聚光鏡U-DCD的光軸上添加一組濾光片和中性密度衰減片,將暗場(chǎng)成像中部分未利用的光源轉(zhuǎn)化為顏色和強(qiáng)度可調(diào)的單色入射光,用于增強(qiáng)具LSPR波長(zhǎng)位于該顏色波段范圍內(nèi)的等離子體顆粒的成像可見(jiàn)度,包括52 nm的AgNPs和長(zhǎng)75.4 nm和直徑38.5 nm的AuNRs,獲得了強(qiáng)于引入的背景光的散射增強(qiáng),同時(shí)保留了較好的成像對(duì)比度。使用軟件的分析結(jié)果與成像效果相符。4.改進(jìn)的成像方法用于小粒徑顆粒和細(xì)胞中顆粒的散射成像。很多情況下,研究對(duì)象的尺寸和空間位阻的問(wèn)題使得納米探針的選擇僅僅限于小粒徑的探針。等離子體納米顆粒的散射能力與半徑的6次方成正比,因此小粒徑的等離子體顆粒的散射強(qiáng)度較弱,這成為制約小粒徑探針廣泛使用的一個(gè)因素。借助上一章設(shè)計(jì)的增強(qiáng)散射成像可見(jiàn)度的設(shè)計(jì),姜黃素包被的20 nm的球形AgNPs以及長(zhǎng)62.6 nm,直徑27.9 nm的AuNPs成像可見(jiàn)度得到了明顯的提升,從成像和RGB線分布數(shù)據(jù)都可以得出以上結(jié)果。復(fù)雜樣品的散射成像容易受到背景的干擾,從而大幅度降低探針的可見(jiàn)度。同樣借助上述的方法,實(shí)現(xiàn)了對(duì)喉癌上皮細(xì)胞中的分散的30 nm藍(lán)色銀納米顆粒和紅色聚集體的高可見(jiàn)度成像。5.雙色等離子體光學(xué)探針對(duì)暗場(chǎng)成像分辨影響的研究�；诘入x子體納米顆粒耦合的納米標(biāo)尺的方法自從被建立以來(lái)就在距離相關(guān)的生化分析得到廣泛應(yīng)用。受限于光學(xué)成像分辨率,近距離的等離子體探針成像時(shí)容易出現(xiàn)光斑的歸并,從而難以從光學(xué)成像上對(duì)單體納米探針的位置進(jìn)行精準(zhǔn)的確定。對(duì)具有不同等離子體散射波段的兩個(gè)納米探針進(jìn)行SEM和暗場(chǎng)散射成像的共定位分析,可以發(fā)現(xiàn),近距離耦合的藍(lán)色AgNP與紅色AgNR在暗場(chǎng)成像光斑中同時(shí)保留了紅色和藍(lán)色,這與相同形體的納米顆粒散射成像結(jié)果是截然不同的。借助這種可以保留各自光學(xué)信息的雙色等離子體納米探針可以用于光學(xué)上成像單體的更佳的分辨。借助窄光源的暗場(chǎng)成像和單色成像重構(gòu)方法可以在一定程度上進(jìn)一步提升紅藍(lán)顆粒的分辨能力�？傊�,本論文借助暗場(chǎng)散射成像技術(shù)實(shí)現(xiàn)了對(duì)納米尺度的反應(yīng)實(shí)時(shí)監(jiān)控以及相關(guān)性質(zhì)的考察和確定。另外通過(guò)改進(jìn)的散射成像方法實(shí)現(xiàn)了對(duì)散射成像可見(jiàn)度的明顯提升,同時(shí)設(shè)計(jì)了通過(guò)散射成像上分辨雙顆粒中單體顆粒位置信息的研究模型,將有利于從等離子體納米探針的散射成像獲取更多的信息。
[Abstract]:Dark field scattering imaging, as a high contrast and non scanning optical imaging technology, is widely used in the fields of sensing, biological process tracer and reaction monitoring. Single nano particle plasma probe has the advantages of the traditional scattering probe, such as stable and strong optical signals and accurate local information. The scattering imaging has a wide application prospect in the field of dynamic response monitoring and spatial resolution in nanoscale. The scattering enhancement and spectrum movement caused by plasma coupling are also widely used in the development of nanoscale scale and biomedical analysis. There are few reports on the reaction process, and there are few reports on the controllable response monitoring of the stimulus response. It may be due to the lack of appropriate research models, imaging signals and dynamic responses that are difficult to establish effective cohesion. It is limited to low utilization of light source and restricted by the constraint of diffraction limit. This paper focuses on the research difficulties and problems mentioned above, drawing on the work basis of predecessors, carrying out specific work in the dynamic response process of the scattered imaging monitoring and the improvement of imaging performance: 1. the matching of the scattering imaging of the dark field Study on the acid sensitivity of nanoscale drug carriers. The coordination polymer microspheres were prepared by the organic ligand 1,1 '- (1,4- Ding Ji) two imidazole and ferrous ions, and the effective in situ encapsulation of adriamycin hydrochloride was realized. The optimal drug loading rate of up to 98% and nearly 40% of the drug could be obtained by the control of the reaction solvent. Load efficiency. The nanoscale drug carrier, due to its own pH sensitivity, and the modification of the cancer cell targeting agent folic acid after the surface silylation, is used for targeted transmission of anticancer drugs and pH sensitive release and controlled-release. The results of fluorescence imaging show that the carrier has a targeting effect and the microsphere morphology of SEM at various time points is studied. The results can prove its acid sensitivity, but can not continuously monitor the single particle level in real time, while the dark field scattering imaging realizes the real-time in situ monitoring of the single particle level of the acid sensitive microsphere carrier degradation process. The acid sensitivity.2. single particle scattering imaging of the drug carrier is directly investigated for the photochemical bond fracture reaction. Real time monitoring of the spherical and rod like silver nanoparticles as the detection probe under the dark field imaging, realized the real-time monitoring of the light sensitivity of the covalent connecting silver - two thiocarbamate (Ag-DTC) chemical bond. By observing the Ag@Ag2S nuclear shell formed by the thiohydrogen root induced by the Ag-DTC fracture The red shift of the scattered signal shows that the cleavage of the chemical bond can be effectively driven by light. At the same time, several other important factors, such as pH, solvent polarity and reducing agent, have also been thoroughly investigated. With the aid of the software analysis of imaging data, the scattering intensity of the spherical AgNPs probe can be accurately found in the process of monitoring the reaction. The single particle scattering spectrum also confirms this phenomenon. For the light driving mechanism, it is explained that photothermal electrons can overcome the energy barrier of the Ag-DTC broken bond and combine the high redox activity of silver, thus showing that the photosensitive.3. that Au-DTC does not have is based on the surface plasmon polariton "light concentration". The effect and the optical path modification is the research of the visibility of the lifting imaging. At present, the dark field imaging mainly uses the oblique illumination, and the excellent imaging contrast is obtained by separating the scattered light from the incident light. In this way, the use efficiency of the light source is low, thus the visibility of the weak scattering signal is damaged, and the power requirement of the light source is also required. The plasma nanoparticles have a scattering cross section that is larger than the physical section, so that the light can be concentrated in the vicinity of the particle, thus scattering a strong light. A group of filters and neutral density attenuation patches are added to the optical axis of the dry dark field spotlight U-DCD, and the light source in the dark field imaging is converted to color and strength. The adjustable monochromatic incident light is used to enhance the imaging visibility of plasma particles with LSPR wavelength in the range of the color band, including 52 nm AgNPs and 75.4 nm and AuNRs with diameter 38.5 nm. The enhanced scattering enhancement of the background light is obtained, and the better imaging contrast is retained. The analysis results and the results of the software are used. In many cases, the size and space hindrance of the object makes the selection of the nano probe limited to the small particle size probe. The scattering ability of the plasma nanoparticles is proportional to the 6 square of the radius, so the small size of the particle size can be obtained in many cases. The weak scattering intensity of plasma particles has become a factor restricting the wide use of small particle diameter probes. With the design of enhanced scattering imaging visibility designed in the last chapter, the visibility of the 20 nm spherical AgNPs of curcumin envelope and the AuNPs imaging of a diameter of 27.9 nm, with a diameter of 27.9 nm, has been significantly improved, from the imaging and RGB line distribution. The above results can be obtained by the data. The scattering imaging of complex samples is easily interfered by the background, thus greatly reducing the visibility of the probe. By the same method, the 30 nm blue silver nanoparticles and the high visibility imaging.5. dual color plasma optical exploration for the laryngeal carcinoma epithelial cells are realized. Research on the influence of dark field imaging resolution. The method of nano scale based on plasma nanoscale coupling has been widely used since it was established. Limited to the resolution of optical imaging, the near distance plasma probe imaging is easy to merge the spot of light, so it is difficult to image from optical imaging. The location of the nanoscale probe is accurately determined. A co localization analysis of the SEM and dark field scattering imaging of two nanoscale probes with different plasma scattering bands can be found. It is found that the near distance coupled blue AgNP and red AgNR retain both red and blue in the dark field imaging spot, which is the same as the same body. The results of scattering imaging of rice particles are completely different. With the use of this kind of dual color plasma nanoscale probe which can retain their respective optical information, the better resolution of optical imaging monomers can be used. The resolution of red and blue particles can be further improved by means of dark field imaging and monochromatic imaging reconstruction with narrow light source. In this paper, the real-time monitoring of the nanoscale reaction and the investigation and determination of the related properties are realized with the help of the dark field scattering imaging technology. In addition, the improved scattering imaging method has been used to improve the visibility of the scattering imaging. At the same time, the location information of the single particles in the double particles is resolved by the scattering imaging. The research model will help to get more information from the scattering imaging of plasma nanoprobes.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：O439
[Abstract]:Dark field scattering imaging, as a high contrast and non scanning optical imaging technology, is widely used in the fields of sensing, biological process tracer and reaction monitoring. Single nano particle plasma probe has the advantages of the traditional scattering probe, such as stable and strong optical signals and accurate local information. The scattering imaging has a wide application prospect in the field of dynamic response monitoring and spatial resolution in nanoscale. The scattering enhancement and spectrum movement caused by plasma coupling are also widely used in the development of nanoscale scale and biomedical analysis. There are few reports on the reaction process, and there are few reports on the controllable response monitoring of the stimulus response. It may be due to the lack of appropriate research models, imaging signals and dynamic responses that are difficult to establish effective cohesion. It is limited to low utilization of light source and restricted by the constraint of diffraction limit. This paper focuses on the research difficulties and problems mentioned above, drawing on the work basis of predecessors, carrying out specific work in the dynamic response process of the scattered imaging monitoring and the improvement of imaging performance: 1. the matching of the scattering imaging of the dark field Study on the acid sensitivity of nanoscale drug carriers. The coordination polymer microspheres were prepared by the organic ligand 1,1 '- (1,4- Ding Ji) two imidazole and ferrous ions, and the effective in situ encapsulation of adriamycin hydrochloride was realized. The optimal drug loading rate of up to 98% and nearly 40% of the drug could be obtained by the control of the reaction solvent. Load efficiency. The nanoscale drug carrier, due to its own pH sensitivity, and the modification of the cancer cell targeting agent folic acid after the surface silylation, is used for targeted transmission of anticancer drugs and pH sensitive release and controlled-release. The results of fluorescence imaging show that the carrier has a targeting effect and the microsphere morphology of SEM at various time points is studied. The results can prove its acid sensitivity, but can not continuously monitor the single particle level in real time, while the dark field scattering imaging realizes the real-time in situ monitoring of the single particle level of the acid sensitive microsphere carrier degradation process. The acid sensitivity.2. single particle scattering imaging of the drug carrier is directly investigated for the photochemical bond fracture reaction. Real time monitoring of the spherical and rod like silver nanoparticles as the detection probe under the dark field imaging, realized the real-time monitoring of the light sensitivity of the covalent connecting silver - two thiocarbamate (Ag-DTC) chemical bond. By observing the Ag@Ag2S nuclear shell formed by the thiohydrogen root induced by the Ag-DTC fracture The red shift of the scattered signal shows that the cleavage of the chemical bond can be effectively driven by light. At the same time, several other important factors, such as pH, solvent polarity and reducing agent, have also been thoroughly investigated. With the aid of the software analysis of imaging data, the scattering intensity of the spherical AgNPs probe can be accurately found in the process of monitoring the reaction. The single particle scattering spectrum also confirms this phenomenon. For the light driving mechanism, it is explained that photothermal electrons can overcome the energy barrier of the Ag-DTC broken bond and combine the high redox activity of silver, thus showing that the photosensitive.3. that Au-DTC does not have is based on the surface plasmon polariton "light concentration". The effect and the optical path modification is the research of the visibility of the lifting imaging. At present, the dark field imaging mainly uses the oblique illumination, and the excellent imaging contrast is obtained by separating the scattered light from the incident light. In this way, the use efficiency of the light source is low, thus the visibility of the weak scattering signal is damaged, and the power requirement of the light source is also required. The plasma nanoparticles have a scattering cross section that is larger than the physical section, so that the light can be concentrated in the vicinity of the particle, thus scattering a strong light. A group of filters and neutral density attenuation patches are added to the optical axis of the dry dark field spotlight U-DCD, and the light source in the dark field imaging is converted to color and strength. The adjustable monochromatic incident light is used to enhance the imaging visibility of plasma particles with LSPR wavelength in the range of the color band, including 52 nm AgNPs and 75.4 nm and AuNRs with diameter 38.5 nm. The enhanced scattering enhancement of the background light is obtained, and the better imaging contrast is retained. The analysis results and the results of the software are used. In many cases, the size and space hindrance of the object makes the selection of the nano probe limited to the small particle size probe. The scattering ability of the plasma nanoparticles is proportional to the 6 square of the radius, so the small size of the particle size can be obtained in many cases. The weak scattering intensity of plasma particles has become a factor restricting the wide use of small particle diameter probes. With the design of enhanced scattering imaging visibility designed in the last chapter, the visibility of the 20 nm spherical AgNPs of curcumin envelope and the AuNPs imaging of a diameter of 27.9 nm, with a diameter of 27.9 nm, has been significantly improved, from the imaging and RGB line distribution. The above results can be obtained by the data. The scattering imaging of complex samples is easily interfered by the background, thus greatly reducing the visibility of the probe. By the same method, the 30 nm blue silver nanoparticles and the high visibility imaging.5. dual color plasma optical exploration for the laryngeal carcinoma epithelial cells are realized. Research on the influence of dark field imaging resolution. The method of nano scale based on plasma nanoscale coupling has been widely used since it was established. Limited to the resolution of optical imaging, the near distance plasma probe imaging is easy to merge the spot of light, so it is difficult to image from optical imaging. The location of the nanoscale probe is accurately determined. A co localization analysis of the SEM and dark field scattering imaging of two nanoscale probes with different plasma scattering bands can be found. It is found that the near distance coupled blue AgNP and red AgNR retain both red and blue in the dark field imaging spot, which is the same as the same body. The results of scattering imaging of rice particles are completely different. With the use of this kind of dual color plasma nanoscale probe which can retain their respective optical information, the better resolution of optical imaging monomers can be used. The resolution of red and blue particles can be further improved by means of dark field imaging and monochromatic imaging reconstruction with narrow light source. In this paper, the real-time monitoring of the nanoscale reaction and the investigation and determination of the related properties are realized with the help of the dark field scattering imaging technology. In addition, the improved scattering imaging method has been used to improve the visibility of the scattering imaging. At the same time, the location information of the single particles in the double particles is resolved by the scattering imaging. The research model will help to get more information from the scattering imaging of plasma nanoprobes.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：O439
【共引文獻(xiàn)】
相關(guān)期刊論文 前10條
1 顏慧瓊;李嘉誠(chéng);馮玉紅;胡文濤;劉若林;林強(qiáng);;改性海藻酸鈉活化SiO_2納米粒子穩(wěn)定載藥Pickering乳液及釋藥性[J];高等學(xué)�；瘜W(xué)學(xué)報(bào);2013年09期
2 韓曉宇;孫德偉;;分子篩作為緩釋藥物載體的研究進(jìn)展[J];廣州化工;2013年18期
3 廖娟;張艷華;葉小舟;廖飛;劉北忠;;氧化石墨烯納米粒與牛血清白蛋白的相互作用[J];光譜實(shí)驗(yàn)室;2013年05期
4 李劍峰;蔣晨;;腦膠質(zhì)瘤靶向藥物遞送系統(tǒng)[J];國(guó)際藥學(xué)研究雜志;2013年05期
5 謝向陽(yáng);林雯;李e，
本文編號(hào)：2124394
本文鏈接：http://sikaile.net/shoufeilunwen/jckxbs/2124394.html
最近更新
教材專(zhuān)著
