1. Thawing: Remove the coating solution from the -20°C freezer and thaw overnight in a 2~8°C refrigerator. After the initial thaw, the product may be aliquoted into small portions for short-term use (e.g., 10–20 mL per aliquot) and stored long-term at -20°C. After use, return the product to the 2~8°C refrigerator promptly. Avoid prolonged exposure to room temperature and repeated freeze-thaw cycles.

2. Coating: Add fully thawed coating solution into TC-treated sterile culture vessels. The recommended dosage is 0.1 mL/cm² (i.e., 1 mL per well of a 6-well plate). Rock the vessel to ensure the coating solution completely covers the bottom surface.

3. Incubation: Place the filled culture vessel in a 37°C incubator and incubate for at least one hour.

4. Storage: If the coated culture vessels are not to be used immediately, they can be kept in a 37°C incubator for 3–5 days, or sealed with sterile parafilm and stored refrigerated at 2~8°C for up to 2 weeks. Warm to room temperature prior to use.

5. Usage: Aspirate the coating solution from the vessel, then immediately add pre-warmed medium or cell suspension for subsequent experiments.