We have decided to measure ROS as an after-thought for my PhD and thus only have the samples that are already frozen. My samples were treated in the following way:
- After animals were euthanised, the brain and quad tissues were snap frozen in liquid nitrogen and stored at -80 degrees C.
- At a later stage, the tissues were very briefly thawed as they were chopped up using a scalpel blade on an ice cold steel table. They were then divided into tubes for all my different analyses, weighed and immediately frozen again on dry ice and then stored at -80.
- The length of time that the samples were thawed for was less than 10 min.
Is there a way to still measure ROS in these tissues?
While temp stress effect on Ros level. But u have control samples when compared nullify the expose but quantitative value slightly vary with fress samples.
Hii Anumesh Pathak,
you can measured the ROS level by using DCFDA assay. You are right that the value will slightly vary with fresh sample but here you can present your value with respect to control. If you have processed the same storage condition for control sample then the induced error in all sample is acceptable as compared to control.