Dear Sophia,

I am no coral expert, but I am not quite sure, if you will detect anything after snap freezing.

ROS are produced and eliminated in every cell in an equilibrium and various ROS sources can be responsible for your production. Since the enzymes and proteins, which normally produce ROS in your cells/tissue are destroyed by freezing, it might be that there will be no ROS to detect. Also the physical influence itself by snap freezing might alter the ROS.

Nevertheless, there is not such a thing as the "ideal general probe for ROS detection". For example, Amplex Red only detects extracellular H2O2. If your cells/tissues produce no extraelllular H2O2 or if the snap-freezing altered H2O2 levels, you might not detect something.

I would like to recommend our review and perspective for further reading concerning ROS probes. If you like please have a look at:

Article Functions of ROS in Macrophages and Antimicrobial Immunity

Article Reactive Oxygen Species: Not Omnipresent but Important in Ma...

For any further questions about ROS measurements feel free to ask any time.

All the best and stay healthy,

Marc

Dear Marc,

Thank-you for that information and those papers. In light of what you have recommended, I believe it may be best to quantify extracellular ROS (H2O2) emitted from fresh tissue cultures taken from coral colonies in our initial experiments. If you have any further recommendations for this please advise. TIA.

All the best,

Sophia

Dear Sophia,

you are very welcome. This sounds like a good experiment to start with. Also isolated coral tissue cells of your choice might be an option.

Please feel free to ask anytime and keep me up to date :)

All the best and stay healthy,

Marc