I used the aging staining kit but the staining was not successful and there were a lot of crystals. What is the reason? My positive control group was not stained successfully either. The x-gal solution had been incubated at 37℃ for more than one hour before use.

1. My positive control sample was treated with etoposide for 24 hours and had been stained before;

2. The pH was not tested. I asked the technician before and the technician said that it was OK to configure it directly;

3. The crystals appeared after staining. The xgal had been incubated for more than one hour before use; The solution did evaporate, and there was more liquid on the lid of the six-well plate.

4. The 37℃ water bath was used for incubation, and a carbon dioxide incubator was not used.