Cryo-FIB lamellae enable ultra-high excitation laser intensities for cryogenic super-resolution fluorescence imaging
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Super-resolution cryogenic correlative light and electron microscopy (SR-cryo-CLEM) integrates molecular specificity and structure on the single-molecule level, but contemporary approaches require low laser powers to preserve specimen vitrification. Here, we perform SR-cryo-CLEM via single molecule localization microscopy (SMLM) on thin biological lamellae and demonstrate that their geometry and minimally-absorbent composition permit excitation laser powers three orders of magnitude higher than standard cryo-EM grids, allowing improved localization precision and imaging speeds.
