Fluo-3AM

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
1. Preparation of Fluo-3 AM working solution
1.1 Preparation of stock solution
Dilute Fluo-3 AM with anhydrous DMSO to prepare a 10 mM stock solution.
Note: It is recommended to aliquot the Fluo-3 AM stock solution and store at -20°C or -80°C protected from light.
1.2 Preparation of working solution
Dilute the stock solution with pre-warmed HBSS to prepare a 1-10 μM Fluo-3 AM working solution.
Note: Adjust the concentration of Fluo-3 AM working solution according to actual needs and prepare fresh before use.
2. Cell staining (suspension cells)
2.1 Collect cells by centrifugation and wash twice with PBS, 5 min each time. Cell density: 1×10⁶/mL.
2.2 Add 1 mL Fluo-3 AM working solution and incubate at room temperature for 5-30 min.
2.3 Centrifuge at 400 g for 3-4 min and discard the supernatant.
2.4 Wash cells twice with PBS, 5 min each time.
2.5 Resuspend cells in 1 mL HBSS and observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslips from the culture medium and aspirate excess medium.
3.3 Add 100 μL dye working solution, gently shake to fully cover the cells, and incubate for 5-30 min.
3.4 Remove the dye working solution, wash 2-3 times with culture medium (5 min each), and observe using a fluorescence microscope.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.