BV-1

Operating Instructions
(The following is a recommended experimental protocol for guidance only; please adjust according to actual requirements)

1. Stock Solution Preparation
1.1 Solvent: Ethanol.
1.2 Recommended concentration: 1 mM.

2. Working Solution Preparation
2.1 Diluent: Serum-free medium.
2.2 Working concentration: 5 μM.
2.3 Notes: The working solution concentration can be adjusted as needed; prepare and use immediately.

3. Staining Procedure
3.1 Sample type: Adherent cells (A549 cells)^[1].
3.2 Incubation conditions: Incubate for 30 min in a humidified environment at 37°C with 5% CO₂.
3.3 Washing step: Wash 3 times with phosphate-buffered saline (PBS) after incubation.

4. Detection and Analysis
4.1 Instrument: Laser scanning confocal microscope; Excitation wavelength (Ex): 488 nm, Emission wavelength (Em): 525-650 nm.
4.2 Result analysis:
4.3 Weak red fluorescence is observable in mitochondria of normal viable cells.
4.4 In cells treated with Monensin or Nystatin, the red fluorescence of mitochondria is significantly enhanced, indicating increased mitochondrial viscosity.
4.5 The fluorescence colocalization result with Mito-Tracker Green confirms its localization in mitochondria, with a Pearson correlation coefficient of 0.98.

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