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Quantification of reactive oxygen species (ROS)

TA Tusar Kanta Acharya
SK Satish Kumar
NT Nikhil Tiwari
AG Arijit Ghosh
AT Ankit Tiwari
SP Subhashis Pal
RM Rakesh Kumar Majhi
AK Ashutosh Kumar
RD Rashmita Das
AS Abhishek Singh
PM Pradip K. Maji
NC Naibedya Chattopadhyay
LG Luna Goswami
CG Chandan Goswami
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Quantification of ROS generation was performed by microscopic analysis using intracellular oxidation of a cell-permeable dye, i.e. 2′, 7′-dichlorodihydrofluorescein diacetate (H2DCFDA) (Invitrogen). The H2DCFDA measures reactive oxygen species (ROS) activity within the cell. Approximately 10,000 bone marrow-derived mesenchymal stem cell population was seeded on glass cover slips and on hydrogel-coated cover slips. Cells were incubated with 10 μm H2DCFDA at 37 °C for 20 min. In a similar manner 50 μm H2O2 (MERCK) treated cells were taken as a positive control. Subsequently the fluorescence signals of ROS were observed under confocal microscope (FV 3000). Images were processed in ImageJ software.

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