We have a molecule tagged with biotin, and then we want to use streptavidin-gold Nanoparticle conjugates to localize it in electron microscopy. Had anybody tried it and has a good experimental procedure, including fixation, incubation times, concentrations, etc.? Moreover, any tips and details we should be careful with?
Gold nanoparticles are too small to see even in electron microscope. Silver amplification is required! The details of the protocol might differ for different subjects.
Immunogold labeling is a standard TEM technique; no problems with visualization of gold nanoparticles. Silver enhancing is possible but not required. Gold is heavy, has excellent contrast against biological tissue.
http://openi.nlm.nih.gov/detailedresult.php?img=2698154_pone.0006084.g009&req=4
I agree that 5 nm or bigger gold particles are well-seen in EM. But the question was about nanogold - ultrasmall particles that are less than 1nm and contain only several Au atoms. I insist that silver amplification is required for those. If somebody managed to detect 1 nanometer gold without amplification, I will greatly appreciate if you send me the publication.
We were looking at large NP, so silver amplificación shouldn't be necessary
Nanoparticle is a particle with at least one dimension less than 100 nm.
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