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.

The above information is based on published literature. Experimental procedures should be appropriately modified to meet specific research demands.