malaria researchers, microbiologists and neuroscientists, how can I detect blood brain barrier breakdown during cerebral malaria in mice other than Evan's blue injection technique?
There are two approaches: The first, if you can't Perfuse your animals and the second if your can. If you cannot perfuse your animals (and can only fix them by submersion in formalin). One approach is to stain for different molecular wt proteins that exist in the sera. These proteins will leak across the BBB. For example you can immunocytochemically stain for IgG. ( While there are a lot of paper that use this approach, note: B cells found in the Brain may produce a confound with this approach, so people also stain for other proteins as well). If you can perfuse, one approach is to follow the Saline perfusion with an HRP solution and then fix en-block with formalin. The you can detect the HRP with a DAB reaction. This is just an explanation to get you started. You can see there are other alternatives to the Evan's blue.
There are two approaches: The first, if you can't Perfuse your animals and the second if your can. If you cannot perfuse your animals (and can only fix them by submersion in formalin). One approach is to stain for different molecular wt proteins that exist in the sera. These proteins will leak across the BBB. For example you can immunocytochemically stain for IgG. ( While there are a lot of paper that use this approach, note: B cells found in the Brain may produce a confound with this approach, so people also stain for other proteins as well). If you can perfuse, one approach is to follow the Saline perfusion with an HRP solution and then fix en-block with formalin. The you can detect the HRP with a DAB reaction. This is just an explanation to get you started. You can see there are other alternatives to the Evan's blue.
It really depends on your experimental design and mouse model. But there are techniques such as IVIS imaging (requires equipment and high cost) or simple injection of FITC-dextran. In any case, I would suggest to use more than one approach.
You can also stain by IHC some key molecules of the BBB integrity and see if they are disrupted. You can do that in addition to other IHC labeling (we use albumin) as Franck said above. We did that for the attached publication (in rats).
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