Upon cellular uptake, the DCFH
-DA probe is hydrolyzed to DCFH
by intracellular esterases and oxidized to highly uorescent dichlorouorescein (DCF) in the presence of ROS.
evaluacion de la capacidadatrapadora de ERO's y ERN's
The ROS release was quantified by determining the increment in DCF fluorescence during imbibition in DCFH
2] because peroxidases, which are reouired for the conversion of DCFH
to DCF, were inhibited by ABAH (Matsugo et al.
Tube C: cells + DCFH
+ PMA (Phorbol MiristateAcetate) - evaluation of chemically induced oxidative burst activity; and
A direct visual investigation was made under an inverted microscope (Optica, 40x and 100x) to observe any morphological changes in the cells cultured with different concentrations of AJLE and/or DCFH
at 24 and 48 h.
In mollusc studies, only DCFH
probe is used to detect the respiratory burst activity.
During this time a part of DCFH
is diffused into the cells.
When ROS exist in the cells, DCFH
can be further oxidized to fluorescent compound DCF, whose fluorescence intensity is proportional to the levels of ROS.
Upon the introduction into the cells, the nonpolar probe DCFH
2-DA was converted into the polar intermediate DCFH
by cellular esterases and further oxidized to highly fluorescent product 2',7'-dichlorofluorescein (DCF) by the intracellular [H.
The fluorescence produced by oxidized DCFH
was measured using a microplate reader at 485 and 528 nm for the excitation and emission wavelengths, respectively.
is converted to fluorescent DCFH
-DA in proportion to the amount of generated intracellular ROS generation.