LysoTracker Blue DND-22

Guidelines (The following is our recommended approach. This guideline is for reference only and should be modified according to your specific needs).
1. Preparation of Stock Solutions and Working Solutions
1.1 Prepare 0.5-1 mM dye stock solutions using anhydrous DMSO.
1.2 Dilute the stock solution with preheated serum-free cell culture medium or PBS to prepare a 50 nM working solution.
Note: Adjust the working solution concentration according to actual conditions, and prepare fresh before use.
2. Cell Staining (Suspension Cells)
2.1 Collect cells by centrifugation, wash twice with PBS, 5 minutes each time.
2.2 Add dye working solution to a final concentration of 50 nM, and incubate at room temperature for approximately 30 minutes.
2.3 Centrifuge at 400 g for 3-4 minutes, and discard the supernatant.
2.4 Wash cells twice with PBS, 5 minutes each time.
2.5 Resuspend cells in serum-free culture medium or PBS, and observe using a fluorescence microscope or flow cytometer. The excitation wavelength is 373 nm, and the emission wavelength is 422 nm to observe lysosomal localization.
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 the dye working solution to a final concentration of 50 nM, gently agitate to completely cover the cells, and incubate for approximately 30 minutes.
3.4 Aspirate the dye working solution, wash 2-3 times with serum-free medium, 5 minutes each time, and observe using a fluorescence microscope or flow cytometry. The excitation wavelength is 373 nm, and the emission wavelength is 422 nm to observe lysosomal localization.
Note: If flow cytometry is required, the cells should be resuspended with trypsin before staining.

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