I am planning to analyse the T-cell population in lymph nodes. Is it okay that I could pool the 2 inguinal lymph nodes after euthanizing the mice and then stain and analyze? Or do I have to do each of the lymph nodes separately?
Generally, these lymph nodes could be pooled. It is also common to see all inguinal, axillary, and brachial LNs pooled and termed cutaneous or skin-draining LN. A circumstance in which pooling these LN would NOT be preferred would be if the two sides of the mouse were treated differently; e.g. one side was sensitized with a chemical while the other side was not. Under these conditions, the cellular composition and/or phenotype of the cells in the LN from the different sides of the mouse may be reasonably expected to be different.
Thanks for the reply. In my case i treat the mouse with one chemical only.
Thanks
Vivek
I always pool inguinal, axillary, and brachial LN before flow analysis. If you have treated the mice with a chemical, maybe you should consider to analyze only the draining LN though.
You can pool as many lymph nodes from each animal provided that you can consistently find them in all your mice. Sometime it happens that you can not find a particular lymph node and this may cause problem. I think depending on your aims of the study you can focus on particular lymph node (s) and always do your analysis on them.
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