New equipment in the RTNN!

The RTNN has several new tools online with many more scheduled for installation in the coming months. Please contact rtnanonetwork@ncsu.edu with any questions regarding technical information or access.

FEI Titan Krios Cryo-Transmission Electron Microscope (Cryo-TEM): This 300 keV instrument offers atomic scale resolution of samples held at cryogenic temperatures and is the most powerful and flexible high-resolution electron microscope for 2D and 3D characterization of biological samples on the market.  Its cryo-based technology and stability allows for single particle analysis and dual-axis cellular tomography of frozen hydrated cell organelles and cells.  The TEM is equipped with a robotic loader, capable of handling up to 12 frozen hydrated samples for increased throughput. Lab renovations are underway for this new Cryo-TEM.

Atomic Force Microscope (AFM): The Asylum AFM includes an MFP-3D head, an XY scanner, and a base. The MFP-3D head offers low noise and precise measurements of the cantilever position for accurate force and topography measurement. The XY-scanner provides flat scans and the ability to accurately zoom and offset with one mouse click. The 3D base offers three configurations for illuminating and viewing samples: top view for opaque samples, bottom view for transparent samples, and dual view for both viewing options.

Raman Microscope: This XploRA PLUS Confocal Raman Microscope includes integrated imaging spectrometer with 4 gratings mounted on motorized turret for full resolution, range and coverage as well as low noise full range CCD detector. It comes with motorized computer controlled 6 position ND filter wheel, confocal pinhole, entrance slit and coupling optics, laser and filter selection. The instrument includes 532 nm and 785 nm Raman excitation laser sources. It offers fast confocal imaging, automated laser wavelength switching. It provides sample identification and chemical imaging on a microscopic scale.

Atomic Layer Deposition (ALD): A newly installed ALD system has allowed us to add 6 precursor lines in one of the facilities, including a low vapor pressure (LVP) delivery line, and has also relieved scheduling pressure that users were experiencing on the original system.  New precursors that we have been exploring since the purchase of the new system include a number of metal organics for depositing CuO, Ga2O3, HfO2, Nb2O5, WO3, and ZnO. The Ultratech Fiji 200Gen 2 Plasma ALD system has four precursor locations and four gas lines into the system for depositions. This instrument is a modular, high-vacuum system that accommodates a wide range of deposition modes.

E-beam lithography: A new Nanometer Pattern Generation System (NPGS) system was installed in one of our focused ion beam (FIB) systems. The NPGS is designed as an e-beam lithography system, but can also be used with the focused ion beam as well.  The system can achieve for patterning with resolution on the order of less than 20 nm for the electron beam and less than 50 nm for the ion beam.

Maskless Lithography System: The Heidelberg Instruments µPG 101 is a direct write lithography system equipped with a 375 nm ultraviolet diode laser capable of exposing feature sizes down to 0.6 µm on either positive or negative photoresists on samples sizes of 10 mm x 10 mm up to 6” x 6”. In addition to full exposure, it has the ability to create surface topographies for gray scale applications.

Reactive ion etcher: The Oxford Plasmalab100 is an induced coupled plasma etcher dedicated to GaN etching.

Cathodoluminescence Imaging and Spectroscopy Detection: The Horiba HCLUE will be installed on our cryo-scanning electron microscope this fall. The spectroscopy system has a focal length of 320 mm and will operate in the ultraviolet-visible range (200-1050 nm). Panchromatic or monochromatic imaging is available in the system.

Retractable Detectors: Two new detectors will be installed on our dual beam FIB/SEM. The retractable annular scanning transmission electron microscope (STEM) detector enables scanning transmission imaging in bright field, dark field, and high-angle dark field modes. The detector includes a special sample holder that can hold up to 6 transmission electron microscopy (TEM) grids and is compatible with the holder used for thin sample preparations. The retractable directional backscatter (DBS) detector features four concentric ring segments that enable separate detection of electrons emitted at different angles. This detector is an ultra-sensitive, Solid State (SS) detector which is sensitive to emitted electrons from 500 V onwards. Using the option beam deceleration, images with beam landing energies down to 50 V are possible. This detector is mounted on a software-controlled retractable arm and allows simultaneous energy dispersive X-ray spectroscopy (EDS) spectra acquisition for WD ≥ 10 mm.

Rapid Thermal Processor: The annealsys AS-One 150 is a rapid thermal processing tool that will be capable of running samples from small pieces to 6” wafers, up to 1300˚C with pressure ranges from atmospheric conditions to high vacuum.

Atomic Force Microscopy (AFM) Workshop

Join us for presentations by academic and industry researchers conducting AFM work. There will also be demonstrations of Bruker’s Dimension Icon with time devoted to test samples. Coffee, tea and donuts will be provided in the morning.
Lunch will be provided at 11:00am for attendees who register.

Schedule:

Technical Talks
9:00 to 11:00 am
Teer 203

“Combined AFM and Vertical Light-Sheet Sideways Microscopy for Living Cell Studies”
Presenter: Dr. Mike Falvo

“New Modes for New Research – From Nanomechanics to Conductivity in Liquid”
Presenter: Dr. Thomas Mueller

Dimension Icon Demos
12:00 to 3:00 pm
Fitzpatrick 1557

Click here for more information, or contact Dr. Mark Walters (mark.walters@duke.edu), Director of SMIF. Register for the workshop here!

Technical Talk Abstracts:

“New AFM modes from Nanomechanics to Conductivity in liquid”
Dr. Thomas Meuller, Director of Applications Development

The Atomic Force Microscope has evolved from a laboratory sensation in the late 80’s to a widely used tool in materials and life science research today. Numerous innovations have driven this evolution, in particular the introduction of new imaging modalities and the parallel development of AFM probe technology, leading to a vast expansion of samples accessible and information revealed by AFM.

In this seminar we will focus on the very advances in Bruker’s AFM technology, including both, novel modes and probes, and the new information they make available enabling new research today. In life sciences, we will show how the evolution of modes from contact to Tapping and PeakForce Tapping has enabled a range of studies from sub molecular structure, where Pyne et al have studied such subtle phenomena as the variability of the DNA double helix structure, to imaging mammalian cells, with first and only AFM images of microvilli on live cells, and extending to cell biomechanical studies that unravel Nanomechanical signatures of cell state, disease progress, and the role of cell membrane receptors. In materials research, we will cover the latest advances in quantitative Nanomechanics of heterogeneous polymer samples and composites at high spatial resolution and at interfaces with PeakForce QNM, FastForce Volume, and contact resonance. Finally, we will cover our most recent extensions of AFM electrical measurements. Having pioneered conductivity measurements on soft and fragile samples with PeakForce TUNA, we have introduced analogous subsurface and impedance measurements with PeakForce sMiM, as well as conductivity in liquid and nanoscale electrochemical measurements with PeakForce SECM.

“Combined AFM and Vertical Light-Sheet Sideways Microscopy for Living Cell Studies”
Mike Falvo, Kellie Beicker, Evan Nelsen, Tim O’Brien, Richard Superfine

The ability of living cells to withstand external mechanical forces and convert mechanical stimuli into biological responses is essential to the physical integrity and biological functions of the cell. Abnormal cellular response to mechanical stimuli has been correlated with disease states, including cancer. Structural information related to rearrangement of the cytoskeletal structure, induced strains, and biochemical distributions are metrics for understanding cell response. However, structural information during applied stress is limited by our ability to image the cells under load, especially in the direction of applied force. In order to study the mechanics of single cells and subcellular components under load, we developed a unique imaging system that combines an atomic force microscope (AFM) with a new imaging technique called vertical light-sheet sideways microscopy (VLSSM). VLSSM combines pathway rotated imaging to facilitate the viewing of downward cell deformation by the applied force of the AFM tip and vertical light-sheet illumination to improve the signal-to-noise ratio achieved by illumination of a thin vertical slice of the cell. The combined AFM-VLSSM system enables simultaneous millisecond imaging and pico-Newton resolution force measurements. We use this new technique to observe force rupture events correlated with detachment of single membrane tethers from a functionalized AFM tip. Force rupture events are correlated with motion of the membrane and intracellular motion, including nuclear deformation attributed to nano-Newton forces applied to attachments at the cell membrane.

Biosensors and Force Measurements in Living Cells

The Chapel Hill Analytical and Nanofabrication Laboratory (CHANL) will host a workshop discussing how and why forces are applied to cells experimentally as well as how responses to these forces are measured using fluorescence-based biosensors, a combined Atomic Force Microscopy, AFM, -optical microscope system combined with vertical light sheet, and magnetic tweezers. In the mornings, participants will hear lectures about various techniques used to apply and measure forces within and generated by cells as well as image analysis tools. In the afternoons, participants will receive hands-on experience with different techniques including AFM, total internal reflection fluorescence (TIRF) microscopy, and magnetic force systems.

More information and a detailed schedule can be found on the event website. Click here to register. Registration fee includes lab supplies, light breakfasts and snacks, and one dinner. Workshop is limited to 18 participants.