Probe system suitable for fluorescence and PET imaging tracing of cells in vivo and preparation method
A fluorescence imaging and cell technology, applied in the field of probe system and preparation, to achieve the effect of easy-to-obtain materials, simple equipment, and strong specificity
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Embodiment 1
[0074] Preparation and characterization of embodiment 1 trans-cyclooctene-PEG-lipid
[0075] (1) trans cyclooctene active ester (in this example, take TCO-PEG4-NHS as an example) and amino lipid (in this example, take DSPE-PEG2000-NH 2 For example) be dissolved in the organic solvent (such as dichloromethane, chloroform or dehydrated alcohol all can) respectively, obtain trans-cyclooctene active ester solution and amino lipid solution respectively, the final concentration of two kinds of solutions is 10mg / mL.
[0076] (2) The trans-cyclooctene active ester solution obtained in step (1) is added to the amino lipid solution, so that the molar number of the trans-cyclooctene active ester is 3 times that of the amino lipid, and the volume ratio is 0.4%. triethylamine, shake gently at room temperature, and react for 8 hours to obtain a reaction solution.
[0077] (3) Remove the organic solvent in the reaction solution of step (2), add an appropriate amount of deionized water to ...
Embodiment 2 4
[0079] Synthesis of embodiment 2 tetrazine near-infrared fluorescent probe (Tz-Cy5)
[0080](1) Tetrazine active esters (Tetrazine-PEG5-NHS Ester is used as an example in this example) and near-infrared fluorescent dyes (Cy5 is used as an example in this example) are dissolved in organic solvents respectively to obtain tetrazine active esters solution and near-infrared fluorescent dye solution.
[0081] (2) The tetrazine active ester solution obtained in step (1) is mixed with the near-infrared fluorescent dye solution, so that the molar number of the tetrazine active ester is 3 times that of the near-infrared fluorescent dye, and the volume ratio is 0.4% triethyl Amines were reacted with gentle shaking at room temperature for 12 h in the dark.
[0082] (3) After the reaction in step (2), use a dialysis bag with a molecular weight cut-off of 1000 Da for dialysis and purification, and freeze-dry to obtain a dark blue powder, which is a tetrazine near-infrared fluorescent probe...
Embodiment 3 4
[0084] Example 3 Preparation and Characterization of Tetrazine-Modified Ultra-Small-Size Water-Soluble Rare Earth Nanoparticle Probes
[0085] (1) Synthesis of oily rare earth nanoparticles NaGdF4-OA modified with oleic acid
[0086] Accurately weigh 0.3717g of gadolinium chloride hexahydrate, add it into a 100mL three-necked flask, and then add accurately measured 4mL of oleic acid and 15mL of octadecene. Under the protection of continuous nitrogen flow, the device was heated to 140°C and magnetically stirred for more than 30 minutes. When all the crystals in the three-necked flask were completely dissolved and a clear solution was formed, the heating was stopped, but nitrogen flow and stirring were maintained until the solution was cooled to room temperature. Separately weigh 0.1 g of solid sodium hydroxide and 0.148 g of ammonium fluoride crystals, sonicate them in 10 mL of methanol solution until they are completely dissolved, then add them dropwise into an Erlenmeyer fla...
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