FM1-43 is a highly lipophilic, water-soluble styrene dye that acts as an osmotic blocker of hair cell mechanical transducer channels and can be used to study neuropeptide granule dynamics and cytotoxicity. dFdCTP trisodium is a Gemcitabine triphosphate trisodium derivative that inhibits DNA synthesis.

Cell staining experiment
a. Solution preparation:
1. Mother solution preparation: Prepare 5 mM mother solution with DMSO, store at -20℃ or -80℃ in the dark after aliquoting.
2. Working solution preparation: Use preheated HBSS solution to prepare 5-20 μM FM 1-43 working solution (select the appropriate working solution concentration according to experimental requirements, and prepare it for immediate use).
b. Cell staining
1. Suspended cell staining
1.1 Collect cells by centrifugation and wash twice with PBS for 5 minutes each time. Add 1 mL FM1-43 working solution and incubate at room temperature for 5-30 minutes.
1.2 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
1.3 Add PBS to wash cells twice for 5 minutes each time.
1.4 Resuspend cells with 1 mL serum-free medium or PBS and observe using a fluorescence microscope or flow cytometer.
2. Adherent cell staining
2.1 Culture iron-adherent cells on sterile slides, discard the culture medium, and remove excess residual liquid.
2.2 Add 100 μL of dye working solution, gently shake to completely cover the cells, and incubate for 5-30 minutes.
2.3 Aspirate the dye working solution, wash with culture medium 2-3 times, 5 minutes each time, and observe using a fluorescence microscope or flow cytometer.