TRITC-DHPE, a rhodamine-labeled glycerophosphoethanolamine lipid, features a brightly red TRITC fluorescence (λEx/λEm=514/580 nm) at its head group. It is utilized for membrane fusion assays and tracing lipid processing in endocytosis. Additionally, TRITC-DHPE serves as an energy transfer acceptor for NBD, BODIPY, and fluorescein lipid probes.

TRITC-DHPE serves as a tool for unilamellar liposome formation and cell labeling. The flow cytometry assay protocolinvolves the following steps: 1. Dissolve the TRITC-DHPE probe in ethanol; 2. Sonicate and dilute with electroporation buffer, sonicate again to reach a final concentration of 4.6 μM; 3. Add 15 mL of the probe stock solution into a T-75 flask and label cells at 37 °C for 2.5 hours; 4. Wash cells twice with PBS, trypsinize, and wash again before analysis; 5. Resuspend cells in Ca 2+ -, Mg 2+ -free PBS and analyze using flow cytometry with 514 nm laser excitation and a 585/42 nm filter; 6. Adjust the aqueous suspension to a pH range of 4-6.5; 7. Create pH-independent emission standard curves, setting the excitation wavelength at 514 nm, emission at 580 nm, with slit widths at 4 nm. For TRITC-DHPE preparation: 1. Store in chloroform (1 mg/ml stock); 2. Dry 1-5 μL of dye stock into a film immediately before use, and reconstitute in 20–100 μL of ethanol; 3. Incubate cells with 100 nM-1 μM final concentration for 5-10 min at 22 °C in RPMI 1640 media with FCS; ensure ethanol concentration does not exceed 1% v/v during incubation.