Table 1.
Probes able to detect specific ROS and RNS species, in vitro, ex vivo or in vivo
| ROS Species | Probe type | Imaging platform used | Tested in vitro | Tested ex vivo/in vivo | ROS/RNS stimulation | Ref |
|---|---|---|---|---|---|---|
| Hypochlorite | Iridium (III) complex-based two-photon phosphorescent probe. | Two-photon laser scanning fluorescence microscope & Confocal laser microscope | HeLa cells/RAW 264.7 cells | Zebrafish | 10mM NaClO (HeLa) 1mg/mL LPS (RAW 264.7 & Zebrafish) |
[201] |
| Hypochlorite | Rhodamine-based hydrazide protein fluorescent probe | Fluorescence microscope | HeLa cells | 50μM OCl− | [200] | |
| Hydroxyl | Ratiometric fluorescence biosensor (gold particles conjugated with organic fluorophore) | Confocal laser microscope | HeLa cells | 10μg/mL LPS | [203] | |
| Hydroxyl | Ratiometric fluorescence biosensor (upconversion nanaoparticles conjugated with organic fluorophore) | Fluorescence microscope equipped with 980nm laser. | HeLa cells | Mouse liver | 500ng/mL PMA (in vitro) 1–4mg LPS/100g body weight (in vivo) |
[202] |
| Superoxide | Fluorescein protein based fluorescent probe | Confocal laser microscope | HCT116/BV-2/RAW 264.7 cells | Zebrafish | 500ng/mL LPS & 50ng/mL IFN-γ (in vitro) PMA 200 ng/mL or antimycin A 500 nM (in vivo) |
[204] |
| Hydrogen Peroxide | Chemo-selective fluorescent naphthylimide peroxide probe | Two-photon laser scanning fluorescence microscope | RAW 264.7 cells | Mouse lung & skin | 1μg/ml LPS (in vitro) 20μg LPS (in vivo) |
[205] |
| Singlet Oxygen | Far-red silicon-rhodamine based chemical fluorescent probe | Fluorescence microscope with 640nm laser | HeLa cells/RAW 264.7 cells | Photosensitizers: 150μg/mL 5-ALA 5μM TMPyP4 |
[206] | |
| RNS Species | ||||||
| Nitric oxide | Chemo-selective copper(II) based fluorescence probe | Confocal laser microscope | HeLa cells/RAW 264.7 cells | 50–200μM DEA-NONOate (HeLa) 200ng/mL LPS (RAW 264.7) |
[208] | |
| Nitric oxide | Far-red two-photon chemical fluorescent probe | Two-photon laser scanning fluorescence microscope | HeLa cells/RAW 264.7 cells | Mouse liver | 25μM NOC-9 (HeLa) 20μg/mL LPS, 200U/mL IFN-γ and 0.5mg/mL L- arginine (RAW 264.7) 1–4mg/ml LPS (in vivo) |
[207] |
| Nitroxyl | FRET-based ratiometric chemical fluorescent probe | Confocal laser microscope | HeLa cells | 100μM AS | [211] | |
| Nitroxyl | Near infra-red chemical fluorescent probe | Confocal laser microscope & In Vivo Imaging System | RAW 264.7 cells | Mouse (in vivo) | 100μM AS (RAW 264.7) 500μM AS (i.p. Mouse) |
[210] |
| Nitroxyl | Lysosome-targetable near infra-red chemical fluorescent probe | Confocal laser microscope & In Vivo Imaging System | RAW 264.7 cells | Mouse (in vivo) | 200μM AS (RAW 264.7) 1mM AS (i.p. Mouse) |
[209] |
| Peroxynitrite | Boronate-based chemical fluorescent probe | Confocal laser microscope | HeLa cells/RAW 264.7 cells | 5 & 20μM Peroxynitrite solution (HeLa) 1μg/mL LPS, 50ng/ml IFN-γ, 2.5ng/ml PMA (RAW 264.7) |
[212] |
LPS: Lipopolysaccharide (produces endogenous ROS/RNS); PMA: phorbol 12-myristate-13-acetate (activates protein kinase C in vivo and in vitro); IFN-γ: Interferon gamma (produces endogenous ROS/RNS); Antimycin A: Produces endogenous ROS/RNS by driving apoptosis; 5-ALA: 5-Aminolevulinic acid (drug used in photodynamic therapy, known to produce singlet oxygen); TMPyP4: 5, 10, 15, 20-tetra-(N-methyl-4-pyridyl)porphyrin (drug used in photodynamic therapy, known to produce singlet oxygen); DEA-NONOate: 2-(N,N-Diethylamino)-diazenolate 2-oxide (Nitric Oxide donor); NOC-9: 6-(2-Hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-hexanamine (Nitric Oxide donor); AS: Angeli’s salt (Nitroxyl donor)