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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.

Achieving the ultimate limits of lithographic resolution and material performance necessitates engineering of matter with atomic, molecular, and mesoscale fidelity. With the advent of scanning helium ion microscopy, maskless He+ and Ne+ beam lithography of 2D materials, such as graphene-based nanoelectronics, is coming to the forefront as a tool for fabrication and surface manipulation. However, the effects of using a Ne focused-ion-beam on the fidelity of structures created out of 2D materials have yet to be explored. Here, we will discuss the use of energetic Ne ions in engineering graphene nanostructures and explore their mechanical, electromechanical and chemical properties using scanning probe microscopy (SPM). By using SPM-based techniques such as band excitation (BE) force modulation microscopy, Kelvin probe force microscopy (KPFM) and Raman spectroscopy, we are able to ascertain changes in the mechanical, electrical and optical properties of Ne+ beam milled graphene nanostructures and surrounding regions. Additionally, we are able to link localized defects around the milled graphene to ion milling parameters such as dwell time and number of beam passes in order to characterize the induced changes in mechanical and electromechanical properties of the graphene surface. [ABSTRACT FROM AUTHOR]

In 1981 A. N. Broers suggested that the spatial limit of direct writing electron beam lithography (DWEBL) would be limited to ∼10 nm by the laterally scattered fast secondary electrons (FSE) even in atomically thin resist. One possible solution to this restriction would be to use low- or ultralow-energy electrons. Experiments and simulations have been carried out to quantify the contribution of FSE to the energy deposition that results in exposure of the resist over high-beam energies. To examine the effects of FSE on low-voltage operations, studies of electron-beam lithography (EBL) in the low- to ultralow-energy range, employing commonly used resists such as PMMA, were performed, and the results were compared to those from conventional high-voltage processing. DWEBL was performed in a Schottky field emission gun scanning electron microscope (SEM), used in cathode-lens mode for ultralow-voltage operation. The exposure characteristics and sensitivity of the system at these energies have been investigated using Monte Carlo simulation methods. Saturation doses were calculated at low energies, which would give a useful condition to target for routine exposure because it ensures the critical dimensions will not be affected by any random changes in beam intensity. [ABSTRACT FROM AUTHOR]