We routinely use the iDISCO+ and Adipoclear techniques followed by imaging on a light sheet system and I find very bright punctate labeling around the outside of tissues such as brain or fat as well as along the inside surfaces of ventricles. We filter our secondary antibodies through a 0.22 um filter as described in the protocol but that does not seem to be enough. Does anyone else have this problem or any suggestions that we can try and reduce it?
Thanks!
David
David Hughes Burk
Thanks, Ra. Unfortunately, that method only uses small molecule dyes for labeling, not antibodies for immunostaining. I also notice little to no background if tissue is stained with dyes such as ToPro, PI, etc.
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