Role of MeCP2, DNA methylation, and HDACs in regulating synapse function

Over the past several years there has been intense effort to delineate the role of epigenetic factors, including methyl-CpG-binding protein 2, histone deacetylases, and DNA methyltransferases, in synaptic function. Studies from our group as well as others have shown that these key epigenetic mechanisms are critical regulators of synapse formation, maturation, as well as function. Although most studies have identified selective deficits in excitatory neurotransmission, the latest work has also uncovered deficits in inhibitory neurotransmission as well. Despite the rapid pace of advances, the exact synaptic mechanisms and gene targets that mediate these effects on neurotransmission remain unclear. Nevertheless, these findings not only open new avenues for understanding neuronal circuit abnormalities associated with neurodevelopmental disorders but also elucidate potential targets for addressing the pathophysiology of several intractable neuropsychiatric disorders

Spatial resolution limits for synchrotron-based infrared spectromicroscopy

Detailed spatial resolution tests were performed on beamline 1.4.4 at the Advanced Light Source synchrotron facility in Berkeley, CA. The high-brightness synchrotron source is coupled at this beamline to a Thermo-Electron Continumum XL infrared microscope. Two types of resolution tests in both the mid-IR (using a KBr beamsplitter and an MCT-A* detector) and in the near-IR (using a CaF2 beamsplitter and an InGaAS detector) were performed and compared to a simple diffraction-limited spot size model. At the shorter wavelengths in the near-IR the experimental results begin to deviate from only diffraction-limited. The entire data set is fit using a combined diffraction-limit and demagnified electron beam source size model. This description experimentally verifies how the physical electron beam size of the synchrotron source demagnified to the sample stage on the endstation begins to dominate the focussed spot size and therefore spatial resolution at higher energies. We discuss how different facilities, beamlines, and microscopes will affect the achievable spatial resolution

Spatial resolution limits for synchrotron-based infrared spectromicroscopy

Detailed spatial resolution tests were performed on beamline 1.4.4 at the Advanced Light Source synchrotron facility in Berkeley, CA. The high-brightness synchrotron source is coupled at this beamline to a Thermo-Electron Continumum XL infrared microscope. Two types of resolution tests in both the mid-IR (using a KBr beamsplitter and an MCT-A* detector) and in the near-IR (using a CaF2 beamsplitter and an InGaAS detector) were performed and compared to a simple diffraction-limited spot size model. At the shorter wavelengths in the near-IR the experimental results begin to deviate from only diffraction-limited. The entire data set is fit using a combined diffraction-limit and demagnified electron beam source size model. This description experimentally verifies how the physical electron beam size of the synchrotron source demagnified to the sample stage on the endstation begins to dominate the focussed spot size and therefore spatial resolution at higher energies. We discuss how different facilities, beamlines, and microscopes will affect the achievable spatial resolution

Temperature measurements with uranyl glass

A preliminary study was completed to determine if uranyl glass could be used as an optical thermometer. Absorption and fluorescence measurements were made as a function of temperature for temperatures ranging from room temperature to approximately 500°C. It was observed that the absorption decreases linearly with increasing temperature. A temperature scale was established for the absorption for the specific setup but it was determined that improvements in the setup need to be made in order to provide measurements that are more useful and repeatable. Each setup needs to be calibrated in order to determine the calibration constants. The fluorescence measurements did not demonstrate a linear relationship to temperature and therefore a temperature scale for fluorescence was not established. It was determined that the absorption is a better method to measure temperature optically using uranyl glass

Spatial resolution limits for synchrotron-based spectromicroscopy in the mid- and near-infrared

Spatial resolution tests were performed on beamline 1.4.4 at the Advanced Light Source in Berkeley, CA, USA, a third-generation synchrotron light source. This beamline couples the high-brightness synchrotron source to a Thermo-Electron Continumum XL infrared microscope. Two types of resolution tests were performed in both the mid-IR and near-IR. The results are compared with a diffraction-limited spot size theory. At shorter near-IR wavelengths the experimental results begin to deviate from diffraction-limited so a combined diffraction-limit and electron-beam-source-size model is employed. This description shows how the physical electron beam size of the synchrotron source begins to dominate the focused spot size at higher energies. The transition from diffraction-limited to electron-beam-size-limited performance is a function of storage-ring parameters and the optical demagnification within the beamline and microscope optics. The discussion includes how different facilities, beamlines and microscopes will affect the achievable spatial resolution. As synchrotron light sources and other next-generation accelerators such as energy-recovery LINACs and free-electron lasers achieve smaller beam emittances, beta-functions and/or energy spreads, diffraction-limited performance can continue to higher-energy beams, perhaps ultimately into the extreme ultraviolet

Spatial resolution limits for synchrotron-based spectromicroscopy in the mid- and near-infrared

Spatial resolution tests were performed on beamline 1.4.4 at the Advanced Light Source in Berkeley, CA, USA, a third-generation synchrotron light source. This beamline couples the high-brightness synchrotron source to a Thermo-Electron Continumum XL infrared microscope. Two types of resolution tests were performed in both the mid-IR and near-IR. The results are compared with a diffraction-limited spot size theory. At shorter near-IR wavelengths the experimental results begin to deviate from diffraction-limited so a combined diffraction-limit and electron-beam-source-size model is employed. This description shows how the physical electron beam size of the synchrotron source begins to dominate the focused spot size at higher energies. The transition from diffraction-limited to electron-beam-size-limited performance is a function of storage-ring parameters and the optical demagnification within the beamline and microscope optics. The discussion includes how different facilities, beamlines and microscopes will affect the achievable spatial resolution. As synchrotron light sources and other next-generation accelerators such as energy-recovery LINACs and free-electron lasers achieve smaller beam emittances, beta-functions and/or energy spreads, diffraction-limited performance can continue to higher-energy beams, perhaps ultimately into the extreme ultraviolet

Abstract Infrared imaging: Synchrotrons vs. arrays, resolution vs. speed

At the ALS we have been testing out Thermo-Electron’s newest infrared imaging system, the Continuum XL microscope. This microscope is equiped with a 32-element MCT (16 · 2) array which allows rapid infrared imaging with fixed step sizes. The microscope also has a conventional single element MCT-A, which can be swapped for an MCT-B, or InSb detectors. This microscope is installed on ALS Beamline 1.4.4 where the synchrotron source provides high brightness for the single element detectors. We present comparisons of the measured spatial resolutions available with each of these detector types for globar and synchrotron sources as a function of wavelength and optical configuration. We find that the synchrotron retains its superiority for ultimate spatial resolution and signal-to-noise, while the array detection system is fast and convenient for surveying larger regions of a sample. Therefore in practice we use the array system for initial infrared images which allow us to find the regions of interest where we ‘zoom in ’ using the synchrotron source