TMA-DPH

TMA-DPH is a hydrophobic fluorescent membrane probe (Ex=355 nm; Em=430 nm). TMA-DPH is able to anchor on the cell surface and localize to different regions of the phospholipid bilayer. By analyzing the fluorescence polarization values of TMA-DPH in the plasma membrane and membrane substructures, the fluidity of the cell membrane can be determined^[1][2][3]. In Vitro:1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 10 mM (optimized according to the experiment).
1.2 Preparation of working solution
Dilute to 0.5-5 μM with PBS or serum-free medium (pH 7.4) (optimized according to the experiment).
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell staining (suspended cells)
2.1 Collect cells by centrifugation and wash twice with PBS for 5 minutes each time.
2.2 TMA-DPH (0.5-5 μM) in Hanks and 20 mM Hepes buffer pH 7.4, and incubate it at 37°C, for 5-30 minutes (optimized according to the experiment).
2.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
2.4 Add PBS to wash the cells twice, 5 minutes each time.
2.5 Wash the cells and resuspended in the appropriate Hepes buffer pH 7.4.
2.6 Observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture the adherent cells on a sterile coverslip.
3.2 Remove the coverslip from the culture medium and remove the excess culture medium.
3.3 Add TMA-DPH (0.5-5 μM) in Hanks and 20 mM Hepes buffer pH 7.4, and incubate it at 37°C, for 5-30 minutes (optimized according to the experiment).
3.4 Wash the cells and resuspended in the appropriate Hepes buffer pH 7.4.
3.5 Fluorescence microscopy detection (Ex/Em = 355/430 nm).
Note: If flow cytometry is required, the cells need to be digested with trypsin and resuspended before staining.