Standardised Measurements for Monitoring and Comparing Multiphoton Microscope Systems.

The goal of this protocol is to enable better characterisation of multiphoton microscopy hardware across a large user base. The scope of this protocol is purposefully limited to focus on hardware, touching on software and data analysis routines only where relevant. The intended audiences are scientists using and building multiphoton microscopes in their laboratories.

Contribution of optical resolution to the spatial precision of two-photon optogenetic photostimulation .

Significance: Two-photon optogenetics combines nonlinear excitation with noninvasive activation of neurons to enable the manipulation of neural circuits with a high degree of spatial precision. Combined with two-photon population calcium imaging, these approaches comprise a flexible platform for all-optical interrogation of neural circuits. However, a multitude of optical and biological factors dictate the exact precision of this approach , where it is most usefully applied.

Propagation of activity through the cortical hierarchy and perception are determined by neural variability.

Brains are composed of anatomically and functionally distinct regions performing specialized tasks, but regions do not operate in isolation. Orchestration of complex behaviors requires communication between brain regions, but how neural dynamics are organized to facilitate reliable transmission is not well understood. Here we studied this process directly by generating neural activity that propagates between brain regions and drives behavior, assessing how neural populations in sensory cortex cooperate to transmit information.

Presynaptic Boutons That Contain Mitochondria Are More Stable.

The addition and removal of presynaptic terminals reconfigures neuronal circuits of the mammalian neocortex, but little is known about how this presynaptic structural plasticity is controlled. Since mitochondria can regulate presynaptic function, we investigated whether the presence of axonal mitochondria relates to the structural plasticity of presynaptic boutons in mouse neocortex. We found that the overall density of axonal mitochondria did not appear to influence the loss and gain of boutons.

Correlative two-photon and serial block face scanning electron microscopy in neuronal tissue using 3D near-infrared branding maps.

Linking cellular structure and function has always been a key goal of microscopy, but obtaining high resolution spatial and temporal information from the same specimen is a fundamental challenge. Two-photon (2P) microscopy allows imaging deep inside intact tissue, bringing great insight into the structural and functional dynamics of cells in their physiological environment. At the nanoscale, the complex ultrastructure of a cell's environment in tissue can be reconstructed in three dimensions (3D) using serial block face scanning electron microscopy (SBF-SEM).

Lamb-dip spectroscopy of the C-N stretching band of methylamine by using frequency-tunable microwave sidebands of CO laser lines.

Lamb-dip spectroscopy of the C-N stretching band of methylamine has been systematically extended to P-, Q-, and R-branch by using microwave sidebands of a large number of CO laser lines as frequency-tunable infrared sources in a sub-Doppler spectrometer. Lamb-dip signals of more than 150 spectral lines have been observed with a resolution of 0.4 MHz and their frequencies have been precisely measured with an accuracy of ±0.

Targeted siRNA Screens Identify ER-to-Mitochondrial Calcium Exchange in Autophagy and Mitophagy Responses in RPE1 Cells.

Autophagy is an important stress response pathway responsible for the removal and recycling of damaged or redundant cytosolic constituents. Mitochondrial damage triggers selective mitochondrial autophagy (mitophagy), mediated by a variety of response factors including the Pink1/Parkin system. Using human retinal pigment epithelial cells stably expressing autophagy and mitophagy reporters, we have conducted parallel screens of regulators of endoplasmic reticulum (ER) and mitochondrial morphology and function contributing to starvation-induced autophagy and damage-induced mitophagy.

Terahertz and far-infrared synchrotron spectroscopy and global modeling of methyl mercaptan, CH3(32)SH.

In this work, terahertz and Fourier transform far-infrared (FTFIR) synchrotron spectra of methyl mercaptan, CH(3)SH, have been investigated in order to provide new laboratory information for enhanced observations of this species in interstellar molecular clouds and star-forming regions. Like its methanol cousin, methyl mercaptan has particularly rich spectra associated with its large-amplitude internal rotation that extend throughout the THz and FIR regions. We have recorded new spectra for CH(3)SH from 1.

High-resolution spectroscopy of the C-N stretching band of methylamine.

The C-N stretching infrared fundamental of CH(3)NH(2) has been investigated by high-resolution laser sideband and Fourier transform synchrotron spectroscopy to explore the energy level structure and to look for possible interactions with high-lying torsional levels of the ground state and other vibrational modes. The spectrum is complicated by two coupled large-amplitude motions in the molecule, the CH(3) torsion and the NH(2) inversion, which lead to rich spectral structure with a wide range of energy level splittings and relative line intensities. Numerous sub-bands have been assigned for K values ranging up to 12 for the stronger a inversion species for the v(t) = 0 torsional state, along with many of the weaker sub-bands of the s species.

Precision Lamb-dip infrared spectra of the C-N stretching band of CH(3)NH(2) with a CO(2)-laser/microwave-sideband spectrometer.

We report saturation dip spectroscopy in the C-N stretching band of CH(3)NH(2) with a resolution of 0.4 MHz and an accuracy of 0.1 MHz by use of a CO(2)-laser/microwave-sideband spectrometer.

Internal rotation and spin conversion of CH3OH in solid para-hydrogen.

The quantum solid para-hydrogen (p-H2) has recently proven useful in matrix isolation spectroscopy. Spectral lines of compounds embedded in this host are unusually narrow, and several species have been reported to rotate in p-H2. We found that a p-H2 matrix inhibits rotation of isolated methanol (CH3OH) but still allows internal rotation about the C-O bond, with splittings of the E/A torsional doublet in internal rotation-coupled vibrational modes that are qualitatively consistent with those for CH3OH in the gaseous phase.

Laser diode photoacoustic detection in the infrared and near infrared spectral ranges.

A new technique for high resolution photoacoustic detection based on application of laser diodes has been developed. This method was tested and compared using identical photoacoustic instrumentation (cell and microphone) to study gas absorption in three different spectral regions, namely: the infrared range near 2100 cm(-1), CO and OCS fundamental band absorption; the ranges near 4200 and 4350 cm(-1), CH4, NH3 and N2O overtone and combination band absorption; the near infrared range near 6500 cm(-1), CO, CO2 and NH3 overtone absorption. Several types of diode laser operating at room temperature or at liquid nitrogen temperature were compared.

Millimeter-Wave Spectra and Global Torsion-Rotation Analysis for the CH(3)OD Isotopomer of Methanol.

New millimeter-wave and microwave measurements for CH(3)OD have been combined with previous literature data and with an extended body of Fourier transform far-infrared observations in a full global analysis of the first two torsional states (v(t) = 0 and 1) of the ground vibrational state. The fitted CH(3)OD data set contained 564 microwave and millimeter-wave lines and 4664 far-infrared lines, representing the most recent available information in the quantum number ranges J View Article and Find Full Text PDF

Dark state illuminated: infrared spectrum and inverted torsional structure of the nu(11) out-of-plane CH3-rocking mode of methanol.

The nu(11) out-of-plane CH3-rocking band of methanol, one of the last fundamentals remaining en route to full vibrational description of this prototype internal rotor, has been identified in the Fourier transform spectrum of CH3OH in the 1150 cm(-1) region. The nu(11) torsional energy pattern is found to be inverted, the first such discovery for a bending state and an important pointer to potential generality of this phenomenon for the whole class of threefold internal rotor molecules. Level-crossing resonances giving transfer channels for intramolecular vibrational energy redistribution (IVR) have been found, and new insights are reported for far-infrared laser emission involving nu(11) levels.

On the Assignment of Optically Pumped Far-Infrared Laser Emission from CH(3)OH.

Progress in the analysis of the infrared spectrum of CH(3)OH in the 930-1450 cm(-1) region has led to assignments, confirmations, or new insights for a number of far-infrared laser (FIRL) transition systems optically pumped by CO(2) lasers. Many of the systems involve FIRL transitions among the CO-stretching, CH(3)-rocking, OH-bending, and CH(3)-deformation vibrational modes, giving useful information on the torsion-rotation structure of the methanol vibrational energy manifold. Some anomalies and mysteries concerning the identity of the lasing levels have been resolved, but several new ones have arisen.

Fourier Transform Infrared Spectroscopy and Vibrational Coupling in the OH-Bending Band of 13CH3OH.

We present in this work a high-resolution Fourier transform infrared study of the OH-bending vibrational band of 13CH3OH. We have investigated the 1070-1400 cm-1 spectral region at 0.002 cm-1 resolution using the modified Bomem DA3.

Fourier Transform Infrared Spectroscopy of the First CO-Stretch Overtone Band of 13CH3OH.

This paper presents a high-resolution Fourier transform infrared study of the first CO-stretch overtone band of 13CH3OH. The spectrum has been recorded at the Justus-Liebig University, Gießsen, Germany on their Bruker IFS 120 HR Fourier transform spectrometer. We have assigned parallel subbands in the torsional state n = 0 for K values up to 6.

C-13 Methanol Far-Infrared Laser: Newly Discovered Lines, Predictions, and Assignments.

We report 25 new laser lines from 13CH3OH methanol when pumped by a cw CO2 laser. The majority of the lines are pumped by the 10R (regular) and 10SR (sequence) bands of the CO2 laser. Two are pumped by the 9HP(20) hot-band CO2 line.

New Measurements and Assignments in the Millimeter-Wave Spectrum of CD3OH.

The ground state rotational spectrum of CD3OH has been revisited in the millimeter-wave range. A total of 216 transition frequencies have been measured and assigned in the 117-179 GHz spectral range, including about 40 transitions previously reported. The spectrum was recorded at the Justus-Liebig University in Giessen, Germany using a frequency modulated millimeter-wave spectrometer.

Global Fit of Torsion-Rotation Transitions in the Ground and First Excited Torsional States of CD3OH Methanol.

A global analysis of reported microwave (MW) millimeter wave (MMW), and Fourier-transform far-infrared (FTFIR) spectra of the CD3OH isotopic species of methanol has been carried out for the first two torsional states (nut = 0 below the barrier and nut = 1 straddling the barrier) of the ground vibrational state. The CD3OH data set contains 472 MW and MMW lines and 5320 FTFIR lines, representing the most recent available information in the quantum number ranges J View Article and Find Full Text PDF

Of what value is a measure of the stutterer's fluency?

The fluency of the stutterer is discussed in terms of how this has contributed to information on stuttering, measurement of efficacy of therapy and possible therapeutic implications. Much research on physiological aspects of stuttering has relied on analysis of fluent tokens and the validity of this is questioned. Use of largely perceptual measures of the quality of fluency after therapy is evaluated and the therapeutic implications of these findings are discussed.