Dear researchers,
I'm planning on quantifying ROS production using DHE superoxide indicator in human macrophages challenged with Mycobacterium tuberculosis. Due to bio-safety issues, samples need to be fixated with PFA 2% overnight prior to flow cytometry analysis. Therefore, I would like to know if anyone had experience using DHE with a fixation step and how it affects the staining.
Also, which other methods could replace PFA fixation and guarantee bacteria killing plus effective DHE staning.
Kindly,
Murilo Delgobo
Hello
kindly look at links here . hope help you
Good Luck
https://www.bdbiosciences.com/documents/webinar_022713_intracellular_flow.pdf
http://mcms.unife.it/it/ricerca-1/ambiti-di-ricerca/patologia-oncologia-e-biologia-sperimentale/signaltransductionlab/publications/143.pdf
Hello
I had used 4%Paraformaldehyde to fixate my cell for 30 minutes，then measured by flow cytometry analysis. maybe you can try.
Good Luck
In context of this topic, I would like to bring to your attention a recent review of our group, "Functions of ROS in macrophages and antimicrobial immunity".
In this review, we give an introduction to ROS and their sources in macrophages, summarize the versatile roles of ROS in direct and indirect antimicrobial immune defense and provide an overview of commonly used ROS probes, ROS source inhibitors and ROS scavengers (also the difference between ROS scavengers and antioxidants, which are not synonymous, is explained).
If you like, please have a look at:
Functions of ROS in Macrophages and Antimicrobial Immunity
February 2021, Antioxidants 10(2):313
DOI: https://doi.org/10.3390/antiox10020313
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