FlavAc

Operating Instructions
Below is our recommended experimental protocol, which serves only as a guide and needs to be adjusted according to your specific requirements.

1. Stock Solution Preparation
1.1 Solvent: DMSO.
1.2 Recommended concentration: 1 mM.

2. Working Solution Preparation
2.3 Diluents: Mouse serum, albumin solution, HDL solution, LDL solution, fetal bovine serum, phosphate-buffered saline (PBS).
2.2 Working concentrations: 1 μM, 5 μM, 10 μM, 50 μM.
2.3 Notes: Adjust the working solution concentration as needed; prepare fresh before use.

3. Staining Procedures
3.1 Sample types^[1]:
3.1.1 In vivo animal samples: Mice.
3.1.2 Ex vivo tissue samples: Isolated mouse aorta, harvested major organs.
3.2 Incubation conditions:
3.2.1 In vivo imaging: Administer intravenously at a dose of 0.6 μmol/kg or 2 mg/kg.

4. Control Setup
4.1 Negative controls: Mice fed a normal diet for fatty liver disease imaging; C57BL/6J mice for atherosclerotic plaque imaging.
4.2 Reference control: Dye solution in DMSO is used for fluorescence retention rate calculation.
4.3 Positive controls are used to validate the experimental system; blank controls are used to eliminate reagent fluorescence interference.

5. Detection and Analysis
5.1 Instruments:
5.1.1 NIR-II fluorescence imaging system with an excitation wavelength of 980 nm.
5.2 Result analysis:
5.2.1 Fluorescence localization: Accumulates in the liver, aortic arch plaques and hindlimb blood vessels of mice.
5.2.2 Color change: NIR-II fluorescence shows cyan/red signals in imaging.
5.2.3 Experimental results: In the fatty liver model, fluorescence intensity increased with increasing duration of high-fat diet; this probe enabled NIR-II imaging of atherosclerotic plaques in ApoE⁻/⁻ mice, with significantly higher fluorescence signals in the plaque region compared to normal vascular tissue.