Design Of Organelles-Targeted Fluorescent Probe For Detection And Imaging Analysis Of Nitric Oxide And Hydrogen Peroxide

Chemo/Biosensing is an important technique to obtain biochemical informations from biological system,such as real time and in situ capture significant informations of biological and pathophysiological processes from living body,living cells and tissues,can be widely used in the field of biomedical analysis,disease diagnosis,etc.Fluorescence imaging techniques involving chemical sensors are essential tools in many fields of science and technology because they are enable to the real-time visualization of informations.For instance,small molecules-based fluorescent probes can attract extensive attention because of its high sensitivity and selectivity.Recently,a lot of research shows detection and imaging of biomolecule in organelles,plays a significant role in study biological mechanism,diagnosis,drug screening,monitoring the response of treatment,etc.Therefore,design and development of targeted organelles fluorescent probes exerts great significance to explore the organelles-related cellular physiological and pathological process.Designing high-efficiency fluorescent probe target to organelles is a hot research domain as well as a difficult problem.Nevertheless,the organelles contain many kinds of active enzyme and organic small molecule probe due to such problems as poor biocompatibility,strong background signal to limit its application.In order to solve these defects efficiently,in this article,in view of the bioactive small molecules,based on the traditional organic small molecule probe,we build a new fluorescent probe platform to meet the organelles analysis and imaging of complex environment.The contents of this paper are as follows:(1)Nitric oxide(NO)is often involved in many different physiological processes including the regulation of lysosomal functions.However,it remains a great challenge to explore the variations of NO levels in lysosomes,limiting the understanding behind its biological functions in cellular signaling pathways and various diseases.Herein,a pH-activatable fluorescent probe,Rhod-H-NO,was designed and synthesized for the determination of lysosomal NO,in which the activation response model is beneficial towards getting accurate biological information.To ensure that Rhod-H-NO can accumulate effectively and exist stably in lysosomes without interference and degradation from other active species,Rhod-H-NO was engineered into the nanopores of mesoporous silica nanoparticles(MSNs)with b-cyclodextrin(?-CD)as the gatekeeper to obtain a nanolab.The nanolab was successfully applied to detect lysosomal NO in living cells and in vivo with high time and spatial resolution.This nanolab could serve as an excellent molecular tool to exploit and elucidate the function of NO at sub-cellular levels.(2)Hydrogen peroxide(H2O2),as a type of reactive oxygen species(ROS),can be endogenously produced from the mitochondrial electron transport chain in aerobic respiration and plays important roles in several physiological processes.Up to date,several two photo probes for cell mitochondrai H2O2 imaging have been designed,but real-time imaging of endogenous H2O2-related biological processes in cell is hampered due to long response time.To address this issue,we report a novel two photo fluorescent probe(CBZ-H2O2)for fast monitoring and imaging H2O2 levels in living cells and tissue.In the presence of H2O2,it exhibits obviously fast response time(about 800s)compared with the reported two photo H2O2 probes.Moreover,the two photo probe CBZ-H2O2 has been successfully applied for the detection and imaging analysis of the intrinsically generated intracellular H2O2 in living cells,and the fluorescence colocalization studies indicate that the probe localizes solely in the mitochondria of HeLa cells.In conclusion,based on conventional organic probe,we constructed dual-control activated fluorescent probe and two photo fluorescent probe for the detection and analysis imaging of biomolecule in subcellular organelles.Achieve real-time and high resolution imaging analysis in specific area of organelles,and overcome defects of strong background signals and susceptible to interference by complex environment which troubled conventional fluorescent probe,affording new ideas and thoughts for design novel fluorescent applied to the detection and analysis in complex systems.