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Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride ([Ru(dpp)3]2+) is an electrochemiluminescent (ECL) probe and oxygen-sensitive sensor. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride can be used to modify electrode surfaces for the detection of sulfates (S2O82-) and oxalates, based on electrochemical reactions that generate excited-state species, releasing photons through irreversible redox reactions. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride utilizes the oxygen quenching of fluorescence mechanism, where fluorescence intensity reflects the metabolic rate of living microorganisms or oxygen levels within cells/tumors, allowing for real-time detection. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride's main applications include microbial detection, antibacterial activity studies, and tumor microenvironment research.

Tris (4,7-diphenyl-1,10-phenanthroline)ruthenium (II) dichloride ([Ru (DPP)3]Cl2) It can be applied to construct a reproducible ECL sensor, which can avoid hydrogen evolution interference when detecting persulfate as a cathode, and at the same time, the anode exhibits a wide linear range in response to oxalate.

1. Experimental steps of fluorescence optical respiration assay (FOR):

1. Will Tris (4,7-diphenyl-1,10-phenanthroline)ruthenium (II) dichloride ([Ru (DPP)3]Cl2) Mixed with silicone rubber, embedded with 2% w/w silicone rubber Add 2 drops of a 96 well plate and incubate in a 37 ℃ wet box for 2-3 days for future use.

2. Cultivate target microorganisms (such as Staphylococcus aureus, Escherichia coli, etc.) in vitro at 37 ℃ for 48 hours under aerobic conditions, and dilute with 10⁵–10⁶CFU/mL; Prepare the experimental drug and dilute it to the set concentration (such as DMSO 2% -40%, medicinal plant extract 0.08 mg/mL -166 mg/mL).

3. Add 80 μL of drug sample or diluted drug formulation (diluted at 1:10, 1:100, 1:1000) to each well of a 96 well plate coated with [Ru (DPP) 3] Cl2, and then add  80 μL of microbial suspension, covering the surface with 60 μL of mineral oil; Use Tecan Infinite 200PRO or other fluorescent enzyme-linked immunosorbent assay (ELISA) reader to monitor the relative fluorescence intensity over time. The experiment is repeated three times, and the time and MIC value at which the half maximum fluorescence intensity is reached are recorded.

4. Using the series dilution method and agar plate colony counting method as controls, verify the consistency of FOR detection results, and evaluate the interference of drug components on the fluorescence signal of [Ru (DPP) 3] Cl2.

2. In vitro experimental steps for tumor oxygen imaging:
1. Preparation of Nano Sensor (Ru ONPS): [Ru (DPP) 3] Cl2 dye was embedded into F-127 polymer and crosslinked with urea and polyformaldehyde to obtain a nano sensor with a particle size of approximately 64 nm.
2. Preparation of cells and co cultured spheroids: Cultivate tumor cells (such as HCT-8 cells) and normal cells (such as NIH3T3 cells) in vitro, and perform routine passaging at logarithmic stages; Construct tumor cell co culture spheres and set the culture time (e.g. 48 h, 120 h, 216 h) for oxygen gradient monitoring.
3. Imaging and oxygen level detection: Ru ONPS is co incubated with tumor cells or co cultured spheres, and the cellular uptake and tumor distribution of the sensor are observed through confocal microscopy; Based on the Stern Volmer equation, oxygen levels were quantitatively detected using changes in sensor fluorescence signals. The detection dynamic range was 0-23 mg/L, with a detection limit of 10 μg/mL. Oxygen content data of cells and co cultured spheres were recorded at different time points (such as 72 hours).

Room temperature in continental US; may vary elsewhere.

2-8°C, protect from light, stored under nitrogen

* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)