Goodafternoon!
I am looking for an easy and quick way to test the BBB integrity after antibiotic treatment: the option now is to image with multiphoton microscope after injection with Dextran. I was wondering if anyone had experience with a quick test in (Lewis) rats.
Thank you!
Depends on your definition of quick and easy, but the Evan's blue assay (aka Miles assay) is certainly more straightforward than multi photon imaging.
Evan's blue binds albumin, so if there is vascular leak the tissue will stain blue. You can compare images or extract the dye and quantify.
See: Article An in vivo Assay to Test Blood Vessel Permeability
Here you can see a comparison (in mice) between this technique and use of radiolabeled antibody.
https://iai.asm.org/content/69/5/3460.short
My thought too. Commonly used in stroke or trauma models, And, Evan's blue is fluorescent too.
I'm assuming the test is done on rodents. If so, why not do an in situ brain perfusion assay using atenolol as an aqueous boundary marker. It would be surprising if the permeability were limited by the flow rate. Maybe the following paper could be useful: A new in situ brain perfusion flow correction method for lipophilic drugs based on the pH-dependent Crone- Renkin equation. Pharm. Res. 2011, 28, 517-530.
I concur with Evans blue as the easiest. There are ways to do in vivo fluorescent imaging now as well using CARS ( Article Chemically-selective imaging of brain structures with CARS microscopy
) . Dyes are always best when available. It is how Erhlich first identified the BBB. If you have PET available, lanthanum can be a good alternative, but is quite expensive. It gives more detail to my knowledge though. Certainly, the correction that Alex mentions using the Crone-Renkin equation is useful if you are looking at permeation.- Badges
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