Wet STEM: A new development in environmental SEM for imaging nano-objects included in a liquid phase

Bogner, Agnès; Thollet, G.; Basset, Dalia M. Abdel; Jouneau, P. H.; Gauthier, Catherine

doi:10.1016/j.ultramic.2005.05.005

Cited by 189 publications

(130 citation statements)

References 11 publications

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“…After applying the twostep QD labeling, and the recording of fluorescence microscopy images, the cells were rinsed with pure water, and imaged in hydrated state using ESEM-STEM 19 . The resulting correlative images provide information about the cellular EGFR distribution, and the stoichiometry in their natural context of hydrated cells.…”

Section: Introductionmentioning

confidence: 99%

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Peckys

Jonge

2015

JoVE

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This protocol describes the labeling of epidermal growth factor receptor (EGFR) on COS7 fibroblast cells, and subsequent correlative fluorescence microscopy and environmental scanning electron microscopy (ESEM) of whole cells in hydrated state. Fluorescent quantum dots (QDs) were coupled to EGFR via a two-step labeling protocol, providing an efficient and specific protein labeling, while avoiding label-induced clustering of the receptor. Fluorescence microscopy provided overview images of the cellular locations of the EGFR. The scanning transmission electron microscopy (STEM) detector was used to detect the QD labels with nanoscale resolution. The resulting correlative images provide data of the cellular EGFR distribution, and the stoichiometry at the single molecular level in the natural context of the hydrated intact cell. ESEM-STEM images revealed the receptor to be present as monomer, as homodimer, and in small clusters. Labeling with two different QDs, i.e., one emitting at 655 nm and at 800 revealed similar characteristic results.

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Section: Introductionmentioning

confidence: 99%

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Peckys

Jonge

2015

JoVE

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“…3 Schematic interactions of engineered nano-pollutants with natural water components humid samples in an ESEM under annular dark-field imaging conditions with resolutions down to a few tens of nm. In this technique, fully submerged samples can be imaged by placing the sample a with TEM grid on a TEM sample holder and placing the whole system inside the ESEM chamber under non-vacuum conditions [11]. Imaging of fully liquid samples is also possible by using AFM which is a member of scanning probe microscopes DLS dynamic light scattering, ESEM environmental scanning electron microscope, FCS fluorescence correlation spectroscopy (confocal microscopy), HPLC high-performance liquid chromatography, Raman Raman spectroscopy, AFM atomic force microscopy, SEM scanning electron microscopy, TEM transmission electron microscopy, UV-Vis UV/Vis spectroscopy, XAS X-ray absorption spectroscopy, XRD X-ray diffraction, XRF X-ray fluorescence spectroscopy, XRM X-ray microscopy, NMR nuclear magnetic resonance a Limit of detection (LOD) (SPMs) family.…”

Section: Microscopy Techniquesmentioning

confidence: 99%

Instrumental approach toward understanding nano-pollutants

Naghdi

Metahni

Ouarda

et al. 2017

Nanotechnol. Environ. Eng.

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Nano-pollutants (NPLTs) have recently raised global concerns due to their possible harmful impact on environment and human health. However, until date, information on the occurrence, fate and toxicity of NPLTs in environment is scant. The knowledge gap can be attributed to the lack of advanced and sophisticated methodologies for the precise detection and characterization of NPLTs at lower concentration in complex matrices, such as surface water, wastewater, soil and food. This review briefly discusses the performance of classical methods for characterization and study of the properties of NPLTs. The important properties include shape, size, aggregation state, chemical composition and structure. Chromatographic, microscopic and spectroscopic techniques have been developed for detection and quantitative estimation of fabricated or naturally existed NPLTs in different matrices. Often, combination of these techniques is required for the separation, purification and accurate estimation. For better detection and understanding of the initial steps of interaction with the environmental matrices, pollution sources, such as wastewater and industrial discharges, must be selected as sampling points. Understanding the dynamics of agglomeration, and decantation will allow to estimate the plume of transport to delimit the potential effects.

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“…Table 1 it is sufficient to obtain information from the thin outer regions of the cells, and those can be imaged with high resolution using environmental scanning electron microscopy (ESEM) with STEM detector (see Figure 2B) (Bogner et al, 2005;Peckys et al, 2013). A third option is given by enwrapping the cells with a graphene sheet (Park et al, 2015;Wojcik et al, 2015).…”

Section: Abbreviationsmentioning

confidence: 99%

Membrane protein stoichiometry studied in intact mammalian cells

Jonge¹

2016

infocus

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Wet STEM: A new development in environmental SEM for imaging nano-objects included in a liquid phase

Cited by 189 publications

References 11 publications

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Instrumental approach toward understanding nano-pollutants

Membrane protein stoichiometry studied in intact mammalian cells

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