CAY10732

Protocol for Imaging of CO in Living Cells^[1]：

Stock Solution Preparation：Stock solutions of CAY10732 (2 mM), PdCl₂ (2 mM), and CORM-3 (10 mM) were separately prepared in HPLC grade DMSO and used freshly. CAY10732 (1 μM) and PdCl₂ (1 μM) in PBS buffer (10 mM, pH 7.4, with 0.5% DMSO, v/v) was prepared in use.

1.HeLa cells were seeded in a 24-well culture plate for one night before cell imaging experiments.

2.In a typical experiment of cell imaging, as controls, living cells were incubated with CAY10732 (1 μM) and a mixture of CAY10732 and PdCl₂ (1 μM each) at 37℃ for 30 min, respectively, and they were imaged after washing with PBS for three times.

3.For imaging of exogenous CO, HeLa cells were pretreated with CORM-3 (1, 5, and 10 μM, respectively) for 30 min at 37℃ and then were incubated with a mixture of CAY10732 and PdCl₂ (1 μM each) for 30 min at 37℃. After washing the cells with PBS buffer, the cells were imaged.

4.For imaging of heme stimulation produced CO, HeLa cells were pretreated with heme (100 μM) for 30 min at 37℃ and then were incubated with a mixture of CAY10732 and PdCl₂ (1 μM each) for 30 min at 37℃. After washing the cells with PBS buffer, the cells were imaged.

This protocol only provides a guideline, and should be modified according to your specific needs.

References:

[1]. Feng, S, Liu, D, et al. Allyl fluorescein ethers as promising fluorescent probes for carbon monoxide imaging in living cells. Anal. Chem. 89(6), 3754-3760 (2017).