Multicolor Electron Microscopy for Simultaneous Visualization of Multiple Molecular Species

摘要: Electron microscopy (EM) remains the primary method for imaging cellular and tissue ultrastructure, although simultaneous localization of multiple specific molecules continues to be a challenge EM. We present obtaining multicolor EM views subcellular components. The uses sequential, localized deposition different lanthanides by photosensitizers, small-molecule probes, or peroxidases. Detailed view biological structures is created overlaying conventional electron micrographs with pseudocolor lanthanide elemental maps derived from distinctive energy-loss spectra each deposit via energy-filtered transmission microscopy. This results in images analogous fluorescence but benefit full spatial resolution illustrate power this methodology visualizing hippocampal astrocytes show that processes two can share single synapse. also polyarginine-based cell-penetrating peptides enter cell endocytosis, newly synthesized PKMζ cultured neurons preferentially localize postsynaptic membrane.