DNA-Lipid探针用于细胞膜外pH值的原位成像
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摘要
将二酰基脂质体自动偶联到两端标有Cy3和Cy5的具有i-motif结构的DNA链上,形成二酰基脂质体-DNA共轭物(DNA-lipid)探针。二酰基脂质体与细胞膜之间强烈的疏水作用可使该探针直接插入细胞膜表面,实现缺氧和常氧条件下细胞外pH值的比率型检测。在高pH值条件下,具有i-motif结构的DNA链两端的荧光基团处于分离状态,无FRET效应;在低pH值条件下,具有i-motif结构的DNA链在细胞膜表面形成四聚体结构,两端的荧光基团相互靠近,产生强的FRET效应。通过测定两种荧光基团的荧光强度比值实现了pH值的定量检测。利用此探针对pH值的灵敏响应实现了对缺氧环境中细胞外pH值的精确测量,在生理病理学上具有重大意义。
Lipid was automatically coupled to DNA chains with i-motif structure marked Cy3 and Cy5 at both ends by DNA synthesizer to form a DNA-lipid probe. The probe could be directly inserted into the surface of cell membrane by strong hydrophobic interaction between lipid and cell membranes,and realized the ratio detection of extracellular pH value under hypoxic and normoxic conditions. Under the condition of high pH,the fluorescent groups at both ends of the DNA strand with imotif structure were separated without any FRET effect,while in the condition of low pH,the DNA strand with i-motif structure formed tetramer structure on the cell surface,and the fluorescent groups at both ends were close to each other,resulting in strong FRET effect. Quantitative detection of pH could be achieved by measuring the fluorescence intensity ratio of the two fluorophores. Accurate detection on extracellular pH in hypoxic environment using this probe is of great significance in physiology and pathology.
Lipid was automatically coupled to DNA chains with i-motif structure marked Cy3 and Cy5 at both ends by DNA synthesizer to form a DNA-lipid probe. The probe could be directly inserted into the surface of cell membrane by strong hydrophobic interaction between lipid and cell membranes,and realized the ratio detection of extracellular pH value under hypoxic and normoxic conditions. Under the condition of high pH,the fluorescent groups at both ends of the DNA strand with imotif structure were separated without any FRET effect,while in the condition of low pH,the DNA strand with i-motif structure formed tetramer structure on the cell surface,and the fluorescent groups at both ends were close to each other,resulting in strong FRET effect. Quantitative detection of pH could be achieved by measuring the fluorescence intensity ratio of the two fluorophores. Accurate detection on extracellular pH in hypoxic environment using this probe is of great significance in physiology and pathology.
引文
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