Rhod-2 AM is a fluorescent probe of the mitochondrion with λex of 552 nm and λem of 581 nm.

Inhibition of mitochondrial Ca2+ uptake by ruthenium red (RR) or DIDS in IMR5 cells infected with poliovirus (PV) is illustrated following fluorescence staining with Rhod-2 AM at 6 h p.i[1].

Detection of Ca2+ changes in mitochondria
I. Solution preparation
1. Mother solution preparation: Dissolve Rhod-2 AM in DMSO to a storage concentration of 10mM.
2. Working solution preparation: Dilute it in serum-free cell culture medium or PBS buffer as needed to obtain a working concentration of 1~10 μM.
II. Operation steps
1. Wash 3 times with serum-free cell culture medium or PBS buffer.
2. Add an appropriate amount of working solution containing Rhod-AM and incubate in a carbon dioxide incubator for 60 minutes. (Adjust according to cell type and dye concentration to ensure that the probe can fully enter the mitochondria)
3. After washing with serum-free cell culture medium or PBS buffer, resuspend the cells in a medium containing serum.
4. Detect by flow cytometry, pay attention to avoid light.