农业工程学报
農業工程學報
농업공정학보
2013年
17期
286-292
,共7页
宋月鹏%康杰%高东升%李江涛%王晓波%尹承苗%毛志泉
宋月鵬%康傑%高東升%李江濤%王曉波%尹承苗%毛誌泉
송월붕%강걸%고동승%리강도%왕효파%윤승묘%모지천
荧光%标记%成像技术%长时程荧光成像%尖孢镰刀菌%活体细胞%碳化硅量子点
熒光%標記%成像技術%長時程熒光成像%尖孢鐮刀菌%活體細胞%碳化硅量子點
형광%표기%성상기술%장시정형광성상%첨포렴도균%활체세포%탄화규양자점
fluorescent%labeling%image technique%long-term-distance fluorescent imaging%Fusarium oxysporum%silicon carbide quantum dots%living cells
致病性尖孢镰刀菌(Fusarium oxysporum)的荧光标记与活体细胞的长时程荧光成像示踪是研究其侵染机制较为有效的方法之一。采用化学腐蚀法制备出 SiC 量子点标记材料,研究了化学腐蚀法制备工艺过程,对 SiC量子点标记材料的微观结构及光学性能进行检测分析,而后对尖孢镰刀菌进行标记及长时程荧光成像,结果表明, SiC 量子点具有良好的生物相容性及光学性能,发射光颜色与其尺寸有内在关联性,腐蚀过程中即可在表面形成多种亲有机物功能团。尖孢镰刀菌生长初期SiC量子点将标记在分生孢子细胞膜处,而后通过内吞作用,进入活体细胞内部并形成稳定标记。进一步研究发现,SiC 量子点无细胞毒性,可以实现活体细胞的长时程荧光成像,同时对其标记特征及原理进行了初步分析讨论。这对于尖孢镰刀菌侵染致病过程组织学与细胞学特征研究,揭示其致病机理、发展新型植株枯萎防治方法提供直观依据和理论分析基础。
緻病性尖孢鐮刀菌(Fusarium oxysporum)的熒光標記與活體細胞的長時程熒光成像示蹤是研究其侵染機製較為有效的方法之一。採用化學腐蝕法製備齣 SiC 量子點標記材料,研究瞭化學腐蝕法製備工藝過程,對 SiC量子點標記材料的微觀結構及光學性能進行檢測分析,而後對尖孢鐮刀菌進行標記及長時程熒光成像,結果錶明, SiC 量子點具有良好的生物相容性及光學性能,髮射光顏色與其呎吋有內在關聯性,腐蝕過程中即可在錶麵形成多種親有機物功能糰。尖孢鐮刀菌生長初期SiC量子點將標記在分生孢子細胞膜處,而後通過內吞作用,進入活體細胞內部併形成穩定標記。進一步研究髮現,SiC 量子點無細胞毒性,可以實現活體細胞的長時程熒光成像,同時對其標記特徵及原理進行瞭初步分析討論。這對于尖孢鐮刀菌侵染緻病過程組織學與細胞學特徵研究,揭示其緻病機理、髮展新型植株枯萎防治方法提供直觀依據和理論分析基礎。
치병성첨포렴도균(Fusarium oxysporum)적형광표기여활체세포적장시정형광성상시종시연구기침염궤제교위유효적방법지일。채용화학부식법제비출 SiC 양자점표기재료,연구료화학부식법제비공예과정,대 SiC양자점표기재료적미관결구급광학성능진행검측분석,이후대첨포렴도균진행표기급장시정형광성상,결과표명, SiC 양자점구유량호적생물상용성급광학성능,발사광안색여기척촌유내재관련성,부식과정중즉가재표면형성다충친유궤물공능단。첨포렴도균생장초기SiC양자점장표기재분생포자세포막처,이후통과내탄작용,진입활체세포내부병형성은정표기。진일보연구발현,SiC 양자점무세포독성,가이실현활체세포적장시정형광성상,동시대기표기특정급원리진행료초보분석토론。저대우첨포렴도균침염치병과정조직학여세포학특정연구,게시기치병궤리、발전신형식주고위방치방법제공직관의거화이론분석기출。
Fluorescence microscopy has allowed the functional study of various molecules that have been identified in living cells. The capabilities of this technique have generated a huge interest in developing new probes for labeling molecules and observing changes in their cellular activities. Quantum dots (QDs) based on II-VI (e.g., CdSe, CdTe, CdS, and ZnSe) and III-V (e.g., InP and InAs) semiconductors have attracted considerable scientific interest over the past two decades for their remarkable luminescent properties. However, the widely used CdX semiconductor QDs were found to be cytotoxic through the release of free metallic ions (cadmium ions for instance). Therefore, it becomes clear that the cytotoxicity strongly influencing biological cell functioning is one of the major limiting factors for the application of II-VI QDs in efficient living cell imaging. Recently, silicon carbide(SiC) QDs has been growing attention for people with the advantages of non-toxic, good biocompatible due to its simple preparation process and excellent optical properties. Especially, for the Fusarium oxysporum, inspection of its infection mechanism needs to be fluorescent marked and long-term-distance tracing with vivo cells. The paper focus on two aspects for SiC QDs: fabrication and the vivo cells marking and long-term-distance fluorescent imaging. <br> Firstly, the aqueous SiC QDs was prepared by SiC particles, which was prepared via self-propagating combustion in the mixture chemical etchant of nitric and hydrofluoric acid. Latticed-hollow of silicon carbide particles will be easily obtained because of SiC particles fabrication characteristics like nonequilibrium thermal forming leading to many defaults on the surface. Processed by broken and dispersed with ultrasonic cavitation, high-speed centrifugation, non-cytotoxic labeling materials of silicon carbide QDs were obtained. The microstructure and optical properties of QDs were measured and analyzed. The silicon carbide QDs are nearly spherical feature and its diameter is shorter than its’ Bohr diameter (5.4 nm), which result in excellent optical properties. There is inherent relationship between the emission color-displaying and the sizes of the SiC QDs. Multi-color fluorescence can be displayed in one exciting wavelength according to different size. Emission light will show blue shift following the decrease diameter. In this study, the emission wavelength of QDs with 5 and 2 nm diameter is 470 and 439 nm, respectively. <br> The Fusarium oxysporum labeling and long-term-distance fluorescent imaging for living cells were processed. Firstly, conidiums were marked on the cell membrance with SiC QDs which next enter into the cell via endocytosis and stably labeling. Further results indicated that SiC QDs can show long-term-distance vivo fluorescent image because of its non-cytotoxicity. <br> Moreover, the results of Fourier transform infrared spectroscopy (FTIR) indicated that some hydrophilic organic functional groups, such as COO-, O-, OH-and so on, on the surface of QDs were established in the process of etching which plays an important role in stable marking for living cells. The problems of interaction between the Fusarium oxysporum and plant root, procedure and mechanism of infecting, new prevention method for Fusarium wilt can be hopefully solved. Furthermore, the SiC QDs is a potential marking materials compared to the traditional fluorescent organic dye. The methods of multi-targets marking and long-term-distance tracing can give the materials and technical support for the biological theory researching fields, such as living cells and its substructure, molecular events, cells behaviors and so on.