RESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMO
Genetically encoded tags for single-molecule imaging in electron microscopy (EM) are long-awaited. Here, we report an approach for directly synthesizing EM-visible gold nanoparticles (AuNPs) on cysteine-rich tags for single-molecule visualization in cells. We first uncovered an auto-nucleation suppression mechanism that allows specific synthesis of AuNPs on isolated tags. Next, we exploited this mechanism to develop approaches for single-molecule detection of proteins in prokaryotic cells and achieved an unprecedented labeling efficiency. We then expanded it to more complicated eukaryotic cells and successfully detected the proteins targeted to various organelles, including the membranes of endoplasmic reticulum (ER) and nuclear envelope, ER lumen, nuclear pores, spindle pole bodies and mitochondrial matrices. We further implemented cysteine-rich tag-antibody fusion proteins as new immuno-EM probes. Thus, our approaches should allow biologists to address a wide range of biological questions at the single-molecule level in cellular ultrastructural contexts.
Assuntos
Ouro/química , Nanopartículas Metálicas/química , Microscopia Eletrônica/métodos , Sistema Livre de Células , Células HeLa , Humanos , Microscopia de Fluorescência , Schizosaccharomyces , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Highly fluorescent bilayer-walled and monolayer-walled nanotubes are assembled from elaborately designed asymmetric perylene diimide (PDI) molecules. The diameter of bilayer-walled nanotubes increases with the size of the branched substituents at the meta-position of the phenyl moiety of PDI molecules, whereas that of monolayer-walled nanotubes remains unchanged regardless of the size of branched substituents.
RESUMO
A series of structurally analogous PDIs were fabricated and used as fluorescent sensor arrays. Adjustment of the molecular electron-donating ability and polarity (i.e., chemical structure) was found to greatly influence the fluorescent quenching by different types of amines. Moreover, the sensor array displayed high sensitivity to amine vapors and allowed the fingerprint differentiation of different species.