Electron microscopy of whole cells in liquid with nanometer resolution

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 cells were placed in buffer solution in a microfluidic device with electron transparent windows insi...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2009-02, Vol.106 (7), p.2159-2164
Main Authors: Jonge, N. de, Peckys, D.B, Kremers, G.J, Piston, D.W
Format: Article
Language:English
Subjects:
Animals
BASIC BIOLOGICAL SCIENCES
Biological Sciences
Buffers
Cell growth
Cells
Cercopithecus aethiops
COS Cells
Electron microscopes
Electrons
Epidermal Growth Factor - metabolism
Equipment Design
FIBROBLASTS
Fibroblasts - metabolism
Fluids
Gold - chemistry
GROWTH FACTORS
Image Processing, Computer-Assisted
Image resolution
Imaging
Liquids
Metal Nanoparticles - chemistry
Microscopy
Microscopy, Electron - instrumentation
Microscopy, Electron - methods
Molecules
Nanoparticles
Nanotechnology
Physical Sciences
Pixels
Protein Binding
Receptor, Epidermal Growth Factor - metabolism
RECEPTORS
SCANNING ELECTRON MICROSCOPY
Silicon
Silicon - chemistry
Silicon nitrides
SKIN
SPATIAL RESOLUTION
TRANSMISSION ELECTRON MICROSCOPY
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Description
Summary: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 cells were placed in buffer solution in a microfluidic device with electron transparent windows inside the vacuum of the electron microscope. A spatial resolution of 4 nm and a pixel dwell time of 20 μs were obtained. The liquid layer was sufficiently thick to contain the cells with a thickness of 7 ± 1 μm. The experimental findings are consistent with a theoretical calculation. Liquid STEM is a unique approach for imaging single molecules in whole cells with significantly improved resolution and imaging speed over existing methods.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0809567106