I am looking to co-localize a phenomenon on DNA via IF and AFM methods. Mica yielded beautiful AFM images, but no IF signal. Glass yielded a great IF image, but was too rough to resolve the DNA on the surface via AFM. What substrate can you recommend that combines both the atomic-flatness of mica with the good optical properties of glass, which would also allow binding of DNA (either with or without some surface treatment)?
Hi, maybe you can use "round glass corverslips" from TedPella. Their roughness is ok for afm measurements.
I would just try to get the mica as thin as possible. Take a scotch tape to remove a thin layer of mica from a bigger sheet. Then try to remove the thin layer from the tape using a tweezer. As soon as you have removed it from the tape you can glue (uv-cureabel adhesive) the thin mica layer to a glass slide.
for instance:
http://www.nature.com/nnano/journal/v9/n3/full/nnano.2013.303.html
Johannes's suggestion is probably the best. Just make sure your glue is clear transparent. Or you can simply stick mica to glass slide with a small amount of ethyleneglycol. This will remove air from the gap between glass and mica and match refractive index throughout your substrate. I am also surprised that mica did not give any signal, I would suggest trying mica of similar thickness to glass slide. It should be transparent enough for fluorescence measurements.
...transparency is one problem with mica, birefringence another, and its surface properties also do not seem perfect for measurements on fluorescence samples. But if you make it thin enough (approx. < 40 micrometer) and mutliply label your DNA (eg. standard intercalators) this should work.
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