I work in the BLT-humanized mouse model with HIV infection, which ravages lymph nodes. In my past, I worked extensively with immune competent mice, which have lovely, large lymph nodes - sadly this is not the case in my model. I am currently training technicians to look for all LN in the hu-BLT mouse +/- HIV infection, but even I have a hard time finding them and then trying to show someone this teeny, tiny pinpoint of slightly different colored ivory is not really helping them. I was wondering if anyone has experience with dyes (I've heard of Chicago sky blue or Evan's blue) for the in vivo detection of lymph nodes such that at sac, we can more easily see the remaining lymph nodes? It would be even better if the dye doesn't effect lymphocyte viability so this could be a routine protocol.
isosulfan blue dye is used clinically to identify "sentinel" lymph nodes. It is injected intradermally and enables visualization of the lymphatic channel and nodes. It should get to the nodes within 5-10 minutes of injection. A single vial is expensive (patient cost $1500) but for mice should be reusable - however, if you are in a hospital that treats melanoma or breast cancer, there should be vials of it each week that have been used, but in which there is a little left over than you could get the operating room to give you for free.
People also have used indocyanine green (ICG) as a near-IR emitter, typically coupled to a colloid particle, which can be visualized through more tissue than the isosulfan blue or india ink and in a thin mouse, should be visible through the skin. It requires an IR source.
Tc99 sulfur colloid is a low energy gamma emitter also used clinically to identify sentinel nodes.
If you look up about these and "sentinel node" you should find references with more details.
I had a similar problem some years ago and visualized the lymph nodes by injection of 100µl 10% (v/v) Indian Ink in isotonic NaCL. This procedure worked very well to visualize the axillary and inguinal lymph nodes and is absolutely non-toxic. However, I have done the injection four days before sac... I do not know whether this procedure works after shorter time periods.
Try using IRDye 800CW PEG from Li-Cor. You will need Odyssey CLX and MousePod or Pearl imager to do this task. I have been using this dye for visualizing inflammation in ears. Let me know if you have any questions. Good luck!
isosulfan blue dye is used clinically to identify "sentinel" lymph nodes. It is injected intradermally and enables visualization of the lymphatic channel and nodes. It should get to the nodes within 5-10 minutes of injection. A single vial is expensive (patient cost $1500) but for mice should be reusable - however, if you are in a hospital that treats melanoma or breast cancer, there should be vials of it each week that have been used, but in which there is a little left over than you could get the operating room to give you for free.
People also have used indocyanine green (ICG) as a near-IR emitter, typically coupled to a colloid particle, which can be visualized through more tissue than the isosulfan blue or india ink and in a thin mouse, should be visible through the skin. It requires an IR source.
Tc99 sulfur colloid is a low energy gamma emitter also used clinically to identify sentinel nodes.
If you look up about these and "sentinel node" you should find references with more details.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2342937/. I like Evans blue. Only caution is that phagocytic cells will pick fluorescence that interferes with flow. Something to do with the dye. I never had any viability issues.
We have used the same india ink trick a Peters above. You can inject iv or ip and find the nodes in about 30 minutes. We have only used this for teaching and have not analyzed cells aftwards, but I suspect they would be fine except the macrophages. Advantage is nothing special is needed for vizualization.
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