Mapping synaptic glutamate transporter dysfunction in vivo to regions surrounding Aß plaques by iGluSnFR two-photon imaging.

Amyloid-ß (Aß) plaques, a hallmark of Alzheimer's disease (AD), are surrounded by regions of neuronal and glial hyperactivity. We use in vivo two-photon and wide-field imaging of the glutamate sensor iGluSnFR to determine whether pathological changes in glutamate dynamics in the immediate vicinity of Aß deposits in APPPS1 transgenic mice could alter neuronal activity in this microenvironment. In regions close to Aß plaques chronic states of high spontaneous glutamate fluctuations are observed and the timing of glutamate responses evoked by sensory stimulation exhibit slower decay rates in two cortical brain areas. GLT-1 expression is reduced around Aß plaques and upregulation of GLT-1 expression and activity by ceftriaxone partially restores glutamate dynamics to values in control regions. We conclude that the toxic microenvironment surrounding Aß plaques results, at least partially, from enhanced glutamate levels and that pharmacologically increasing GLT-1 expression and activity may be a new target for early therapeutic intervention.

High Q optical fiber tips for NC-AFM in liquid.

Non-contact atomic force microscopy is rapidly expanding from ultra-high vacuum to include the study of surfaces and biomolecules in liquids by high resolution imaging and force spectroscopy. This is despite the additional frequency shift noise due to the inherently low Q factor of the cantilever oscillating in a liquid. In this paper we present a tip based on an optical fiber which can operate in liquid with Q factors in excess of 100 using a 'diving bell' arrangement which allows only a small portion of the tip to be submerged. We demonstrate stable imaging and force spectroscopy using this set-up. The tips are based on scanning near-field optical microscopy tips and, when used with NC-AFM, provide a method of combining both high resolution mechanical and fluorescence studies of biomolecules and cells.

Determination of the local contact potential difference of PTCDA on NaCl: a comparison of techniques.

There has been increasing focus on the use of Kelvin probe force microscopy (KPFM) for the determination of local electronic structure in recent years, especially in systems where other methods, such as scanning tunnelling microscopy/spectroscopy, may be intractable. We have examined three methods for determining the local apparent contact potential difference (CPD): frequency modulation KPFM (FM-KPFM), amplitude modulation KPFM (AM-KPFM), and frequency shift-bias spectroscopy, on a test system of 3,4,9,10-perylene tetracarboxylic dianhydride (PTCDA) on NaCl, an example of an organic semiconductor on a bulk insulating substrate. We will discuss the influence of the bias modulation on the apparent CPD measurement by FM-KPFM compared to the DC-bias spectroscopy method, and provide a comparison of AM-KPFM, AM-slope detection KPFM and FM-KPFM imaging resolution and accuracy. We will also discuss the distance dependence of the CPD as measured by FM-KPFM for both the PTCDA organic deposit and the NaCl substrate.