Carboxyfluorescein modified kidney clearance type silver nanoparticles, and preparation method and application thereof
A carboxyfluorescein, silver nanoparticle technology, applied in nanotechnology, nanotechnology, nanotechnology and other directions for materials and surface science, can solve the problems of photobleaching emission lifetime, high toxicity, low absorption coefficient, etc., to reduce Effects of photobleaching, small organism toxicity, high processability
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[0027] The preparation method of the above-mentioned carboxyfluorescein-modified kidney-scavenging silver nanoparticles comprises the following steps:
[0028] (1) Mix 97mL deionized water, 2mL, 100mmol / L silver nitrate solution, 1mL, 200mmol / L glutathione solution, add 5mol / L sodium hydroxide solution to adjust the pH to 4.9-5.1, and then mix Add 200 μL of sodium borohydride solution with a concentration of 0.5mol / L to the solution, and vigorously stir at a stirring speed of not less than 900 r / min for 2 days at room temperature;
[0029] (2) After the above solution was reacted for 2 days, the reaction was stopped, the reaction solution was taken out and transferred to a dialysis bag, dialyzed for 3 days, and finally freeze-dried for 24 hours to obtain GS-AgNPs powder;
[0030](3) Redissolve the above GS-AgNPs powder in PBS to prepare a 0.05mmol / L solution, adjust the pH to 8-9, and mix the pH-sensitive fluorescent dye 5(6)-carboxyfluorescein N-succinimide The ester was dis...
Embodiment 1
[0033] Fluorescence, ultraviolet absorption, HR-TEM, electrophoresis and other tests were carried out on a kind of ultra-stable silver nanoparticles FG-AgNPs prepared by the present invention.
[0034] The fluorescent dye NHS-FAM was coated with silver nanoparticles GS-AgNPs, and FG-AgNPs were synthesized by the preparation method of the above-mentioned examples. figure 1 This is the HR-TEM image of the synthesized FG-AgNPs. It can be seen that the particles have a spherical structure, good dispersion, and a size with a diameter of less than 5.5nm meets the requirements for renal clearance in organisms.
[0035] FG-AgNPs prepared as in the present invention, figure 2 (a) The absorption spectra of GS-AgNPs, FG-AgNPs, and NHS-FAM are given, as shown in the figure below, NHS-FAM has a strong absorption peak at 490 nm. GS-AgNPs did not show any significant uptake. exist figure 2 In (b), the fluorescence spectrum of NHS-FAM shows a maximum wavelength at 550 nm, while FG-AgNPs ...
Embodiment 2
[0037] Take 10 μL of the FG-AgNPs prepared by the present invention and add them into the small sample groove of the rubber plate, connect the wires for electrophoresis, stop the electrophoresis when it moves to 1-2 cm from the front, and observe under the ultraviolet light after taking it out. image 3 (a) is the gel-electrophoresis picture of FG-AgNPs and GS-AgNPs prepared in the present invention and the dye molecule NHS-FAM. Lane 1 represents GS-AgNPs, light yellow in lane 2 shows FG-AgNPs formed by NHS-FAM conjugated with GS-AgNPs, dark yellow in lane 3 represents free NHS-FAM dye. Although all 3 lanes move in the same direction, GS-AgNPs and FG-AgNPs move at the same speed and at the same time as seen from the free NHS-FAM dye. Furthermore, comparison of the bands in lanes 2 and 3 shows that unreacted dye was successfully removed by the Sephadex technique. Binding was further confirmed by fluorescence images of agarose gels visualized under UV light, as image 3 (b) sh...
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