Correlative 3D superresolution fluorescence and electron microscopy reveal the relationship of mitochondrial nucleoids to membranes

摘要: Microscopic images of specific proteins in their cellular context yield important insights into biological processes and architecture. The advent superresolution optical microscopy techniques provides the possibility to augment EM with nanometer-resolution fluorescence access precise location ultrastructure. Unfortunately, efforts combine have been stymied by divergent incompatible sample preparation protocols two methods. Here, we describe a protocol that preserves both delicate photoactivatable fluorescent protein labels essential for fine ultrastructural EM. This enables direct 3D imaging 500- 750-nm sections interferometric localization followed scanning generated focused ion beam ablation. We use this process “colorize” detailed mitochondrion position labeled proteins. approach presented here has provided new level definition vivo nature organization mitochondrial nucleoids, expect straightforward method be applicable many other questions can answered imaging.