A new rhodamine B derivative T1 has been rationally synthesized and displayed selective Pd（Ⅱ）-amplified absorbance and fluorescence emission above 540 nm in methanol-water. Upon the addition of Pd（Ⅱ）, the spirol...A new rhodamine B derivative T1 has been rationally synthesized and displayed selective Pd（Ⅱ）-amplified absorbance and fluorescence emission above 540 nm in methanol-water. Upon the addition of Pd（Ⅱ）, the spirolactam ring was unfolded and a 1：1 metal-ligand complex formed, which can be used for ＇＇naked-eyes＂ detection. In addition, fluorescence imaging experiments of Pd^（2＋） in HepG2 living cells showed its valuable application in biological systems.展开更多
Studying the expression level of mRNA in living cells will offer tremendous opportunities for ad-vancement in cell biology research,disease diagnostics,and drug discovery.In this paper,a molecular beacon(MB)specific f...Studying the expression level of mRNA in living cells will offer tremendous opportunities for ad-vancement in cell biology research,disease diagnostics,and drug discovery.In this paper,a molecular beacon(MB)specific for the important tumor suppressor gene p21 has been designed and synthesized.The fluorescence signal was detected in real-time after the MB entered the cytoplasm of nasopharyn-geal carcinoma cells.After injecting the p21MB into nasopharyngeal carcinoma cell and p33-trans-fected nasopharyngeal carcinoma cell,the consistent increase of fluorescent signal intensity was de-tected in both cell lines,and maximum fluorescence intensity achieved in about 15 min.In about 4 min following microinjection,the fluorescence increasing rate was significantly different between these two cell lines,which indicate the different p21 mRNA expression levels.The results obtained in the real-time detection were also validated by RT-PCR.Analysis of the initial fluorescence increasing rate can effi-ciently reduce the side effect of enzyme and improve the accuracy in living cell mRNA detection.展开更多
In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate...In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection.展开更多
扫描离子电导显微镜(scanning ion conductance microscopy,SICM)是一种非接触式的扫描探针显微技术(scanning probe microscopy,SPM),可以实现生物样品在近生理条件下的成像。随着技术发展,目前广泛应用于生物医学领域的SICM主要包括两...扫描离子电导显微镜(scanning ion conductance microscopy,SICM)是一种非接触式的扫描探针显微技术(scanning probe microscopy,SPM),可以实现生物样品在近生理条件下的成像。随着技术发展,目前广泛应用于生物医学领域的SICM主要包括两种:跳跃式离子电导显微技术(hopping probe ion conductance microscopy,HPICM)和外加压力模式的SICM。前者可以应用于软的、黏的、对外力或其它机械信号敏感的样品的高分辨成像;后者可以通过探针微管对样品局部施加外力刺激或化学、电学、光学或生物分子等信号,实现对样品动力学性质或相关生理过程局部的原位研究。此外,SICM技术具有良好的开放性,能够越来越多地与其它技术手段联用,极大地丰富了其在生物医学领域的应用,可用于疾病发病机理、药物作用以及临床诊断等的研究。但是,目前SICM时间分辨率较低,这制约了它在生物体系动力学行为方面的研究。展开更多
基金supported by the State Key Laboratory of Analytical Chemistry for Life Science,Jiangsu Key Laboratory of Advanced Organic Materials,School of Chemistry and Chemical Engineering,Nanjing University。
基金supported by the National Natural Science Foundation of China (No. 21172178)
文摘A new rhodamine B derivative T1 has been rationally synthesized and displayed selective Pd（Ⅱ）-amplified absorbance and fluorescence emission above 540 nm in methanol-water. Upon the addition of Pd（Ⅱ）, the spirolactam ring was unfolded and a 1：1 metal-ligand complex formed, which can be used for ＇＇naked-eyes＂ detection. In addition, fluorescence imaging experiments of Pd^（2＋） in HepG2 living cells showed its valuable application in biological systems.
基金the National Key Basic Research Program(Grant No.2002CB513110)the Key Technologies Research Development Program of China(Grant Nos.2003BA310A16and2005EP090026)+2 种基金International Technologies Collaboration Program of China(Grant No.2003DF000039)the National Natural Science Foun-dation of China(Grant Nos.90606003and20475015)Key Project of Hunan Province Technology Plan of China(Grant No.0399Y1006)
文摘Studying the expression level of mRNA in living cells will offer tremendous opportunities for ad-vancement in cell biology research,disease diagnostics,and drug discovery.In this paper,a molecular beacon(MB)specific for the important tumor suppressor gene p21 has been designed and synthesized.The fluorescence signal was detected in real-time after the MB entered the cytoplasm of nasopharyn-geal carcinoma cells.After injecting the p21MB into nasopharyngeal carcinoma cell and p33-trans-fected nasopharyngeal carcinoma cell,the consistent increase of fluorescent signal intensity was de-tected in both cell lines,and maximum fluorescence intensity achieved in about 15 min.In about 4 min following microinjection,the fluorescence increasing rate was significantly different between these two cell lines,which indicate the different p21 mRNA expression levels.The results obtained in the real-time detection were also validated by RT-PCR.Analysis of the initial fluorescence increasing rate can effi-ciently reduce the side effect of enzyme and improve the accuracy in living cell mRNA detection.
基金Supported by the National Key Basic Research Program (Grant No. 2002CB513110)International Technologies Collaboration Program of China (Grant No. 2003DF000039)National Natural Science Foundation of China (Grant Nos. 90606003 and 20620120107)
文摘In this study, GFP mRNA in COS-7 cell and GFP-transfected COS-7 cell was detected in real time using phosphorothioate-modified molecular beacon based on living cell imaging method. Results showed that phosphorothioate-modified molecular beacon still kept the advantages of molecular beacon, such as, excellent selectivity, high sensitivity, and no separation detection. In addition, this modification could significantly increase the nuclease resistance of molecular beacon. Phosphorothioate-modified molecular beacon can efficiently reduce the false positive signal and improve the accuracy of living cell mRNA detection.
文摘扫描离子电导显微镜(scanning ion conductance microscopy,SICM)是一种非接触式的扫描探针显微技术(scanning probe microscopy,SPM),可以实现生物样品在近生理条件下的成像。随着技术发展,目前广泛应用于生物医学领域的SICM主要包括两种:跳跃式离子电导显微技术(hopping probe ion conductance microscopy,HPICM)和外加压力模式的SICM。前者可以应用于软的、黏的、对外力或其它机械信号敏感的样品的高分辨成像;后者可以通过探针微管对样品局部施加外力刺激或化学、电学、光学或生物分子等信号,实现对样品动力学性质或相关生理过程局部的原位研究。此外,SICM技术具有良好的开放性,能够越来越多地与其它技术手段联用,极大地丰富了其在生物医学领域的应用,可用于疾病发病机理、药物作用以及临床诊断等的研究。但是,目前SICM时间分辨率较低,这制约了它在生物体系动力学行为方面的研究。
