Fluorescence Microscopic Image Analysis of Nucleic Acids Based on The Capillary Flow Directed Assemb

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It is critical to establish a direct and precise method with a high sensitivity and selectivity in analytical chemistry. In this research, making use of a well known phenomenon of capillary flow, we have proposed an image analysis method of nucleic acids at the price of a small amount of sample. When a droplet of the supramolecular complex solution, formed by neutral red and nucleic acids(NA) under an approximate neutral condition, was placed on the hydrophobic surface of dimethyl dichlorosilane pretreated glass slides, and it was evaporated, the supramolecular complex exhibited the periphery of the droplet due to the capillary effect, and accumulated there to form a red capillary flow directed assembly ring(CFDAR). A typical CFDAR has an outer diameter of (2 r ) about 1.18 mm and a ring width(2 δ ) of about 41 μm. Depending on the experimental conditions, a variety of CFDAR can be assembled. The experimental results are in agreement with our former theoretical discussion. It was found that when a droplet volume is 0.1 μL, the fluorescence intensity of the CFDAR formed by the NR NA is in proportion to the content of calf thymus DNA in the range of 0-0.28 ng, fish sperm DNA of 0-0.24 ng and yeast RNA of 0-0.16 ng with the limit of detection(3 σ ) of 1 7, 1.4 and 0.9 pg, respectively for the three nucleic acids. It is critical to establish a direct and precise method with a high sensitivity and selectivity in analytical chemistry. In this research, making use of a well aware phenomenon of capillary flow, we have proposed an image analysis method of nucleic acids at the price of a When a droplet of the supramolecular complex solution, formed by neutral red and nucleic acids (NA) under an approximate neutral condition, was placed on the hydrophobic surface of dimethyl dichlorosilane pretreated glass slides, and it was evaporated, the supramolecular complex exhibited the periphery of the droplet due to the capillary effect, and accumulated there to form a red capillary flow directed assembly ring (CFDAR). A typical CFDAR has an outer diameter of (2 r) about 1.18 mm and a ring width δ) of about 41 μm. Depending on the experimental conditions, a variety of CFDAR can be assembled. The experimental results are in agreement with our former theoretical discussion . The was found that when a droplet volume is 0.1 μL, the fluorescence intensity of the CFDAR formed by the NR NA is in proportion to the content of calf thymus DNA in the range of 0-0.28 ng, fish sperm DNA of 0-0.24 ng and yeast RNA of 0-0.16 ng with the limit of detection (3σ) of 1 7, 1.4 and 0.9 pg, respectively for the three nucleic acids.
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