1. Thawing: Remove the VTN from a -80°C to -20°C freezer and allow it to thaw slowly in a 2°C to 8°C refrigerator, or thaw it at room temperature and immediately return it to a 2°C to 8°C refrigerator. After the initial thaw, the product may be aliquoted into single-use portions (50–200 μL per aliquot) for long-term storage at -80°C to -20°C. After use, return the product to a 2–8°C refrigerator as soon as possible to avoid prolonged exposure to room temperature and repeated freeze-thaw cycles.

2. Preparation of the working solution: Dilute VTN with Ca²⁺- and Mg²⁺-free DPBS at a ratio of 1:100, and mix thoroughly by pipetting.

3. Coating: Add the prepared working solution to a sterile culture dish treated with TC. The recommended volume is 0.1 mL/cm² (i.e., 1 mL per well in a six-well plate). Gently shake the dish to ensure the working solution completely covers the bottom surface.

4. Incubation: Incubate the culture vessels containing the added solution in a 37°C incubator for at least one hour.

5. Storage: If the coated culture vessels are not used immediately, they may be stored in a 37°C incubator for 3–5 days, or sealed with sterile sealing film and refrigerated at 2–8°C for up to 2 weeks. Allow them to return to room temperature before use.

6. Usage: Remove the VTN working solution from the container and immediately add it to pre-warmed culture medium or cell suspension for subsequent experiments.