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Table 2 Available probes for detecting ROS in C. elegans

From: Caenorhabditis elegans as a powerful tool in natural product bioactivity research

Probe

Reaction

Target

Pros

Cons

Refs

2’,7’-Dichlorofluorescein diacetate

Oxidation of 2′,7′-dichlorodihydrofluorescein to fluorescent 2’-7’-dichlorofluorescein

H2O2, •HO, and ROO•

Detects ROS in the whole body

- Exerts free radical properties itself

- Lacks specificity

[31]

Amplex red

Oxidation of 10-acetyl-3,7-dihydroxypenoxazine by horseradish peroxidase in the presence of H2O2 that produces a red fluorescence

H2O2

- Both fluorometric and spectrophotometric measurement

- Detect ROS in the surrounding medium

- Causes autooxidation and a self-amplification of the fluorescent signal

- Forms superoxide in the presence of NADH

[154, 155]

MitoSOX

Oxidation of dihydroethidium by O2• − to fluorescent ethidium

Mainly O2• − 

- Targets the mitochondria

- Provides less background fluorescence signal

- Possibly reacts with cell components like cytochrome C

[31]

MitoTracker Red CM-H2Xros

Oxidation of the reduced form to the red-fluorescent rosamine

H2O2

Mitochondria selectivity

- Poor detection of other ROS

- Background signal generated from nonspecific lipid uptake of dye

[33]

Nuclear Peroxy Emerald 1

H2O2-mediated conversion of arylboronates to phenol

H2O2

Measuring nuclear H2O2 fluxes

 

[39]

Peroxy orange I

Oxidation of the fluorophore masked by boronate moiety by H2O2 to unmasked fluorescence compound

H2O2

 

- Competes with H2O2-scavenging enzymes

- Reacts with ONOO–

[35]

MCLA

Forms N-methylacridone upon oxidation by O2• − 

O2• − and singlet oxygen

Does not undergo redox cycling

- Low specificity

- High background noise

[31, 36]

CellROX

Bright green fluorescence upon oxidation by ROS and subsequent binding to mitochondrial DNA

O2• − and •HO

Detects many types of ROS

 

[34, 37]

CBH

Oxidizes the benzeneboronic group to phenol after reacting with H2O2

H2O2

High sensitivity and selectivity to H2O2

 

[38]

DCHP

DCHP is oxidized to a fluorescent dicyanomethylene-4H-pyran derivative (DCM)

H2O2

High sensitivity and selectivity to H2O2

 

[40]