Laurdan (solution)

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Preparation of working solution^[1]
The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
2. Laurdan staining
2.1 Cells are cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 5 mL Pen-Strep, and 2.5 mL 1 M HEPES.
2.2 Freshly isolated cells are seeded into 35 mm Mattek glass-bottom dishes coated with fibronectin.
2.3 Cells are incubated with Laurdan for 40 min at 37°C.
2.4 Observe using an LSM710 META laser scanning microscope.
2.5 SimFCS software processing to calibrate the system and phasor diagram of FLIM data by measuring fluorescein (pH 11).

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