Nanoscale imaging of whole cells using a liquid enclosure and a scanning transmission electron microscope.
Nanoscale imaging techniques are needed to investigate cellular function at the level of individual proteins and to study the interaction of nanomaterials with biological systems. We imaged whole fixed cells in liquid state with a scanning transmission electron microscope (STEM) using a micrometer-sized liquid enclosure with electron transparent windows providing a wet specimen environment. Wet-STEM images were obtained of fixed E. coli bacteria labeled with gold nanoparticles attached to surface membrane proteins. Mammalian cells (COS7) were incubated with gold-tagged epidermal growth factor and fixed. STEM imaging of these cells resulted in a resolution of 3 nm for the gold nanoparticles. The wet-STEM method has several advantages over conventional imaging techniques. Most important is the capability to image whole fixed cells in a wet environment with nanometer resolution, which can be used, e.g., to map individual protein distributions in/on whole cells. The sample preparation is compatible with that used for fluorescent microscopy on fixed cells for experiments involving nanoparticles. Thirdly, the system is rather simple and involves only minimal new equipment in an electron microscopy (EM) laboratory.
Peckys
Diana B
DB
Center for Environmental Biotechnology, University of Tennessee, Knoxville, Tennessee, United States of America.
Veith
Gabriel M
GM
Joy
David C
DC
de Jonge
Niels
N
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D066246:ErbB Receptors / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000648:ultrastructure; D006046:Gold; D046208:Microchip Analytical Procedures; D017348:Microscopy, Electron, Scanning Transmission / Q000295:instrumentation*; D057054:Molecular Imaging / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D010316:Particle Size; D014867:Water
D014867:Water; D006046:Gold; D066246:ErbB Receptors