Advice on preserving mouse cervical lymph nodes for flow cytometry?
Hi everyone,
I'm optimizing a flow cytometry protocol using mouse cervical lymph nodes from a disease model. Our goal is to identify the immune cell populations present and determine which cells are associated with dextran-coated iron nanoparticles injected into the mice.
As a test, we used snap-frozen lymph nodes, but after dissociation the cells were almost all dead and the staining was very poor. I'm trying to figure out whether snap-freezing is simply not suitable for preserving viable immune cells or if something else may have gone wrong.
Ideally, I'd like to preserve the tissue for a few weeks before processing.
Does anyone have recommendations for the best way to do this while maintaining good cell viability?
Would you freeze the intact lymph nodes or prepare a single-cell suspension first? Budget-friendly options (e.g., homemade cryopreservation media) would be especially appreciated.
Side question: We also need to detect the dextran-coated iron nanoparticles by flow. The only reagent I've found so far is an FITC-conjugated anti-dextran antibody. If anyone has experience detecting these nanoparticles or has alternative approaches, I'd love to hear your suggestions.
Thanks in advance!