Application of Second Harmonic Imaging Combined with Fluorescence Imaging for Locating Transdermal Absorption of Substances
A substance and transdermal technology, applied in the field of biomedicine, can solve the problems of inability to dynamically observe the position and retention of drugs entering the skin, and inability to mark the position information of macromolecular drugs, and achieve the effect of simple operation and clear characterization.
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[0027] Example 1. Detection of Human Collagen Transdermal Absorption by Second Harmonic Combined with Two-Photon Fluorescence Imaging
[0028] 1. Fluorescently labeled R-hc
[0029] Accurately weigh 100 mg of human collagen R-hc dry powder, add 10 mL of phosphate buffer to fully dissolve it, and use 1mol / L NaHCO 3 The solution was adjusted to pH 8.5±0.5. iFluor TM Add 1ml of anhydrous DMSO to 350 (1mg) to obtain a 1mg / mL dye solution. While stirring with a magnetic stirrer, add the fluorescent dye solution drop by drop to the protein solution to avoid sticking the fluorescein to the wall of the flask (the addition should be completed within about 20 minutes). After the addition, continue to stir in the dark for about 12 hours. During the binding period, the protein solution should be kept at about 4°C, so the beaker and stirrer should be moved together into a 4°C refrigerator. Take out the fluorescently labeled protein and put it into a dialysis bag, put it into a large be...
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