Single gold-tagged epidermal growth factor (EGF) molecules bound to cellular EGF receptors of fixed fibroblast cells were imaged in liquid with a scanning transmission electron microscope (STEM). The placed buffer solution microfluidic device transparent windows inside the vacuum microscope. A spatial resolution 4 nm and pixel dwell time 20 micros obtained. layer was sufficiently thick contain thickness 7 +/- 1 microm. experimental findings are consistent theoretical calculation. Liquid STEM...

Abstract Monte Carlo softwares are widely used to understand the capabilities of electron microscopes. To study more realistic applications with complex samples, 3D needed. In this article, development version CASINO is presented. The software feature a graphical user interface, an efficient (in relation simulation time and memory use) model, accurate physic models for microscopy applications, it available freely scientific community at website: www.gel.usherbrooke.ca/casino/index.html . It...

The intracellular uptake of 30 nm diameter gold nanoparticles (Au-NPs) was studied at the nanoscale in pristine eukaryotic cells. Live COS-7 cells were maintained a microfluidic chamber and imaged using scanning transmission electron microscopy. A quantitative image analysis showed that Au-NPs bound to membranes vesicles, possibly lysosomes, occupied 67% available surface area. vesicles accumulated form micrometer-sized cluster after 24 h incubation. Two clusters analyzed found consist 117 ±...

We present a microfluidic system that maintains liquid flow in specimen chamber for scanning transmission electron microscope (STEM) imaging. The consists of two ultrathin silicon nitride windows supported by microchips. They are placed holder seals the sample from vacuum and incorporates tubing to connected syringe pump outside microscope. Using results obtained fluorescence microscopy microspheres flowing through system, an equation characterize was calibrated. Gold nanoparticles diameters...

The imaging of microscopic structures at nanometre-scale spatial resolution in a liquid environment is interest for wide range studies. Recently, flow transmission electron microscopy (TEM) holder equipped with microfluidic cell has been developed and shown to exhibit nanoparticles through an transparent viewing window. Here we demonstrate the application system both scanning conventional immobilized few nanometres water micrometre thickness. TEM bright field be limited by chromatic...

Gold nanoparticles were observed to move at a liquid/solid interface 3 orders of magnitude slower than expected for the movement in bulk liquid by Brownian motion. The nanoscale was studied with scanning transmission electron microscopy (STEM) using enclosure consisting microchips silicon nitride windows. experiments involved variation dose, coating nanoparticles, surface charge enclosing membrane, viscosity, and thickness. slow not result hydrodynamic hindrance near wall but instead...

A graphene liquid cell for transmission electron microscopy (TEM) uses one or two sheets to separate the from vacuum in microscope. In principle, is an excellent material such application because it allows highest possible spatial resolution, provides a flexible covering foil, and effectively protects evaporating. Examples open literature have demonstrated atomic-resolution TEM using small pockets coverage of whole biological cells with sheets. total three different basic types are...

The formation of HER2 homodimers plays an important role in breast cancer aggressiveness and progression; however, little is known about its localization. We have studied the intra- intercellular variation at single-molecule level intact SKBR3 cells. Whole cells were visualized hydrated state with correlative fluorescence microscopy environmental scanning electron (ESEM). locations individual receptors detected using anti-HER2 affibody combination a quantum dot (QD), fluorescent...

Liquid-phase electron microscopy (LPEM) is capable of imaging native (unstained) protein structure in liquid, but the achievable spatial resolution limited by radiation damage. This damaging effect more pronounced when targeting small molecular features than for larger structures. The matter even complicated because critical dose that a sample can endure before damage not only varies between proteins also critically depends on experimental conditions. Here, we examined beam observed...

Electron emission measurements were conducted on individual carbon nanotubes. The nanotubes had a closed end and their surfaces thoroughly cleaned. It is shown conclusively that nanotube electron emitters indeed exhibit Fowler–Nordheim behavior have work function of 5.1±0.1eV for the under investigation, which diameters 1.4 4.9nm.

Silicon microchips with thin, electron transparent silicon nitride windows provide a sample support that accommodates both light-, and microscopy of whole eukaryotic cells in vacuum or liquid, minimum preparation steps. The are robust enough cellular samples can be cultured directly onto them, no addition supporting film, there is need to embed section the sample, as typically required microscopy. By combining two microchips, microfluidic chamber constructed for imaging liquid microscope. We...

Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and study interaction nanomaterials with biological systems. We imaged whole fixed cells in liquid state a scanning transmission electron microscope (STEM) using micrometer-sized enclosure transparent windows providing wet specimen environment. Wet-STEM images were obtained E. coli bacteria labeled gold nanoparticles attached surface membrane proteins. Mammalian (COS7) incubated...

Correlative fluorescence microscopy and transmission electron (TEM) is a state-of-the-art methodology to study cellular function, combining the functionality of light with high resolution microscopy. However, this technique involves complex sample preparation procedures due its need for either thin sections or frozen samples TEM imaging. Here, we introduce novel correlative approach capable imaging whole eukaryotic cells in liquid scanning (STEM); there no additional necessary Quantum dots...

Electron microscopy of biological cells in liquid provides unique nanoscale information. A highly attractive idea is the capability to also study physiological processes live with electron microscopy. However, this seems unrealistic because minimal needed dose obtain contrast already many orders magnitude above lethal known cause reproductive-cell death. We show here that claims viable recent reports are based on a questionable interpretation used fluorescence live/dead assay. practical...

We demonstrate electron microscopy of fully hydrated eukaryotic cells with nanometer resolution. Living Schizosaccaromyces pombe were loaded in a microfluidic chamber and imaged liquid scanning transmission (STEM). The native intracellular (ultra)structures wild-type three different mutants studied without prior labeling, fixation, or staining. STEM images revealed various components that identified on the basis their shape, size, location, mass density. maximal achieved spatial resolution...