Lyso-VR1

Operating Instructions
(The following is a recommended experimental protocol for guidance only, and adjustments are required based on your specific needs).

1. Working Solution Preparation
1.1 Diluent: PBS.
1.2 Working concentration: 1 μM.

2. Staining Procedure
2.1 Sample type: Adherent cells (HeLa cells)^[1]
2.2 Incubation conditions: Incubate with 1.0 μM Lyso-VR1 for 30 min.
2.3 Washing step: Wash once with PBS after incubation.

3. Control Setup
3.1 Set up unstained negative control cells.
3.2 Set up control cells cultured in normal medium.

4. Detection and Analysis
4.1 Instruments: Confocal fluorescence microscope; Flow cytometer.
4.1.1 Excitation wavelength: 488 nm; Emission wavelength: 500 nm-580 nm.
4.2 Result analysis:
4.2.1 The fluorescence intensity increases with the elevation of lysosomal viscosity, which is particularly evident during starvation-induced mitophagy.
4.2.2 Fluorescence localization: Specific targeting of lysosomes.
4.2.3 Fluorescence color: Green.