How to store the immunostained sample after imaging?

xyang

Hello,

I am starting to try CLARITY and immunostaining on my 1-mm-thick mouse brain slices. I am wondering how do people store their immunostained samples after imaging in case they are needed for a second time imaging sometime later? I am concerned that if storing in PBS or clearing solution will let antibodies diffuse out over time and if that means the sample can only be imaged within a short period of time after immnostaining? Are there any tricks to prevent antibodies from leaking out of the sample?

Thanks!

yglasson

Hello,
I store my samples in PBS-azide0.1%. I can image them 1 month later without loosing of the signal!

xyang

Thanks a lot for your info!

ClearLightBio

Samples should always be washed and stored in PBS after the initial imaging since most, if not all RI match solutions, are not storage solutions. If it is for long-term storage it needs to be in PBS with sodium azide. As long as the sample remains hydrated you should be able to return to it. We’ve successfully imaged tissues that we cleared 2 years prior. Samples should be covered if they have been immunostained and you wish to reimage them at a later date. You may need to restain your tissue though if it has been a significant amount of time since the initial staining as the signal might fade. That is the nature of the fluorophores conjugated to the fluorophores. We always recommend a fresh counterstain on the sample after prolonged storage.