I would like to intradermally inject FITC or TRITC labelled dextran conjugates into the dorsal site of the moue ear and intradermally into the side flank. I would like to either use these stained vessels for intravital 2P-LSM or fix the ear and side flank 4%PFA and then use it for imaging. I only want to stain the lymph tics, so i.v. injection is no option.
In the Literature usually 40kDa-2000kDa dextrans are used, but very often the concentration and the time until the dextran dye is taken up is not indicated.
I already tried it with 70kDa FITC-dextran and waited once 30 min and another time 2h before invivo 2P imaging but the dye was not taken up by the lymphatics - rather randomly distributed in the tissue, no vessels structures were detectable at all (imaging away from the injection site of the dye).
Evans blue dye, which is commonly used for visualisation has a molecular weight of 900Da and works really nice- so I wonder why in so many publications 40-2000kDa of dextran conjugates are used for lymphatics.
So now my questions:
What is the optimal MW of dextran-conjugates for staining lymphatic vessels of the ear and side flank?
How long do I have to wait until the dye is taken up efficiently so that I can image the vessels?
Is intradermal injection the right method?
Thanks
Agnes
Dear Agnes,
I have no personal experience with this model, but based on the literature, I would recommend you to Dex-150.
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0057135
http://paperity.org/p/33638825/optimization-and-regeneration-kinetics-of-lymphatic-specific-photodynamic-therapy-in-the
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