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 inside the vacuum of the electron microscope. A spatial resolution of 4 nm and a pixel dwell time of 20 micros were obtained. The liquid layer was sufficiently thick to contain the cells with a thickness of 7 +/- 1 microm. 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.
de Jonge
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Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, TN 37232-0615, USA. niels.de.jonge@vanderbilt.edu
Peckys
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Kremers
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Piston
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D000818:Animals; D002021:Buffers; D019556:COS Cells; D002522:Chlorocebus aethiops; D004815:Epidermal Growth Factor / Q000378:metabolism; D004867:Equipment Design; D066246:ErbB Receptors / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D053768:Metal Nanoparticles / Q000737:chemistry; D008854:Microscopy, Electron / Q000295:instrumentation* / Q000379:methods*; D036103:Nanotechnology; D011485:Protein Binding; D012825:Silicon / Q000737:chemistry
D002021:Buffers; D004815:Epidermal Growth Factor; D006046:Gold; D066246:ErbB Receptors; D012825:Silicon