Frequently Asked Questions

If you are looking to try out Magnify for yourself, there are a few good places to start:

• For those entirely new to ExM procedures, protocol papers from older methods can get you started on the basics. Be on the lookout for something Magnify-specific soon as well!
• Head over to our Protocols page, where you can find the most up-to-date protocols, reagents, and tips.
• Please feel free to contact us with any questions that you still have and we will be happy to help!

For larger endeavors, please contact us as well and we would be happy to discuss a collaborative project.

There sure are! Head on over to our Protocols page for a full list.

For many of these, they are just what we use and have found to work well. An important exception is sodium acrylate (SA), one of the main monomers in the Magnify gel. Expansion success can vary wildly based on SA quality. We recommend either purchasing from Santa Cruz or AK Scientific, but most strongly recommend Santa Cruz.

Additionally, it is imperative to note SA quality (even from the proper vendor) before use in a monomer solution. SA should be a fine white powder. If it is yellowed or clumpy, or there is any undissolved debris floating in the monomer solution, it is probably best to retry with something more fresh. It is normal for the monomer solution to have a yellow tint, however.

Probably! Please feel free to visit our validated protocols page, where we outline some gel and homogenization conditions for common tissues including many FFPE human clinical samples, PFA-fixed mouse brain, and human lung organoid. If your tissue of interest is not included here, there is still a high likelihood that Magnify will be compatible with it. A good place to start is to try one of our validated protocols and optimize from there. Alternatively, you may contact our lab and we would be happy to assist you. We are continually refining our approach and any newly validated tissue types will be added to the website.

Also probably! We have had a high rate of success with antibodies for post-expansion staining (~80%). We even find that often, antibodies will work very well with Magnify that don’t work in unexpanded tissue, because the homogenization process functions as a highly effective antigen retrieval protocol.

In our hands, it is rare (though not impossible) for an antibody that works well in unexpanded tissue to stop working well after Magnify processing. However, if a normally reliable antibody isn’t giving expected results post-expansion, there are a couple of things you can try to troubleshoot:

• Insufficient washing of the tissue after homogenization is the most common cause of antibody failure in Magnify samples. SDS from the homogenization solution must be thoroughly washed out. Our favorite method is by using C12E10 non-ionic surfactant, washing 3 x 10 minutes (or more) at room temperature, followed by 60 minutes at 60C in C12E10, before a final 3 x 10 minute wash at room temperature.
• Since expanded tissues are much thicker than unexpanded tissue, it can take a little longer to perform blocking than normal. In our hands, leaving your expanded tissue in blocking solution (e.g. 3% BSA in 0.1% Triton X-100 / 1x PBS) for an hour is generally sufficient to ensure complete blocking.

Magnify takes a similar amount of time to other ExM methods. Tissue gelling can be done either overnight, or can be completed in as few as four hours, with only a small percentage of this time being hands-on. Homogenization with hot surfactant varies by tissue type. Cultured HEK cells, for instance, can be completely homogenized in 6 hours. Some tissue types and fixation methods take longer, e.g. PFA-fixed mouse brain (8 hours for 30-50 µm sections), FFPE human brain (10 hours), and FFPE human kidney (60 hours). Antibody staining protocols take the same amount of time as in unexpanded tissues.

Yes! The Magnify gel is compatible with Proteinase K digestion, and the Magnify homogenization procedure is compatible with any ExM gel, at least for softer tissues such as PFA-fixed mouse brain or human lung organoid.

However, for best results it is highly recommended to follow the complete Magnify protocol. We have had great success recovering fluorescent protein signal with anti-FP primary antibodies, and the Magnify gel gives distortion-free expansion with the broadest range of samples.