Two-photon imaging with diffractive optical elements

Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE) which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopies

Cognitive and Physiologic Impacts of the Infraslow Oscillation

Brain states are traditionally recognized via sleep-wake cycles, but modern neuroscience is beginning to identify many sub-states within these larger arousal types. Multiple lines of converging evidence now point to the infraslow oscillation (ISO) as a mediator of brain sub-states, with impacts ranging from the creation of resting state networks (RSNs) in awake subjects to interruptions in neural activity during sleep. This review will explore first the basic characteristics of the ISO in human subjects before reviewing findings in sleep and in animals. Networks of consistently correlated brain regions known as RSNs seen in human functional neuroimaging studies oscillate together at infraslow frequencies. The infraslow rhythm subdivides nonREM in a manner that may correlate with plasticity. The mechanism of this oscillation may be found in the thalamus and may ultimately come from glial cells. Finally, I review the functional impacts of ISOs on brain phenomena ranging from higher frequency oscillations, to brain networks, to information representation and cognitive performance. ISOs represent a relatively understudied phenomenon with wide effects on the brain and behavior

Temporal coupling of field potentials and action potentials in the neocortex

The local field potential (LFP) is an aggregate measure of group neuronal activity and is often correlated with the action potentials of single neurons. In recent years, investigators have found that action potential firing rates increase during elevations in power high‐frequency band oscillations (50–200 Hz range). However, action potentials also contribute to the LFP signal itself, making the spike–LFP relationship complex. Here, we examine the relationship between spike rates and LFP in varying frequency bands in rat neocortical recordings. We find that 50–180 Hz oscillations correlate most consistently with high firing rates, but that other LFP bands also carry information relating to spiking, including in some cases anti‐correlations. Relatedly, we find that spiking itself and electromyographic activity contribute to LFP power in these bands. The relationship between spike rates and LFP power varies between brain states and between individual cells. Finally, we create an improved oscillation‐based predictor of action potential activity by specifically utilizing information from across the entire recorded frequency spectrum of LFP. The findings illustrate both caveats and improvements to be taken into account in attempts to infer spiking activity from LFP.We examined the relationship between spike rates and local field potentials (LFP) in the rat neocortex, and we find that while 50–180 Hz oscillatory power correlates most consistently with firing rates of neurons, other LFP bands also carry spiking‐related information. We additionally find that spiking itself and electromyographic activity contribute to LFP power and that the ratio of excitatory to inhibitory activity also correlates with 50–180 Hz power. Finally, we create an improved oscillation‐based predictor of action potential activity by utilizing information from the entire LFP frequency spectrum at once.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/1/ejn13807-sup-0001-FigS1.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/2/ejn13807-sup-0007-FigS7.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/3/ejn13807-sup-0002-FigS2.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/4/ejn13807-sup-0003-FigS3.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/5/ejn13807-sup-0005-FigS5.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/6/ejn13807_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/7/ejn13807-sup-0009-reviewerComments.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/8/ejn13807-sup-0006-FigS6.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/9/ejn13807-sup-0008-FigS8.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/10/ejn13807.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146325/11/ejn13807-sup-0004-FigS4.pd

SLM Microscopy: Scanless Two-Photon Imaging and Photostimulation with Spatial Light Modulators

Laser microscopy has generally poor temporal resolution, caused by the serial scanning of each pixel. This is a significant problem for imaging or optically manipulating neural circuits, since neuronal activity is fast. To help surmount this limitation, we have developed a “scanless” microscope that does not contain mechanically moving parts. This microscope uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern. This allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision. To demonstrate the usefulness of this microscope, we perform two-photon uncaging of glutamate to activate dendritic spines and cortical neurons in brain slices. We also use it to carry out fast (60 Hz) two-photon calcium imaging of action potentials in neuronal populations. Thus, SLM microscopy appears to be a powerful tool for imaging and optically manipulating neurons and neuronal circuits. Moreover, the use of SLMs expands the flexibility of laser microscopy, as it can substitute traditional simple fixed lenses with any calculated lens function

Two-Photon Microscopy with Diffractive Optical Elements and Spatial Light Modulators

Two-photon microscopy is often performed at slow frame rates due to the need to serially scan all points in a field of view with a single laser beam. To overcome this problem, we have developed two optical methods that split and multiplex a laser beam across the sample. In the first method a diffractive optical element (DOE) generates a fixed number of beamlets that are scanned in parallel resulting in a corresponding increase in speed or in signal-to-noise ratio in time-lapse measurements. The second method uses a computer-controlled spatial light modulator (SLM) to generate any arbitrary spatio-temporal light pattern. With an SLM one can image or photostimulate any predefined region of the image such as neurons or dendritic spines. In addition, SLMs can be used to mimic a large number of optical transfer functions including light path corrections as adaptive optics

UP States Protect Ongoing Cortical Activity from Thalamic Inputs

Cortical neurons in vitro and in vivo fluctuate spontaneously between two stable membrane potentials: a depolarized UP state and a hyperpolarized DOWN state. UP states temporally correspond with multineuronal firing sequences which may be important for information processing. To examine how thalamic inputs interact with ongoing cortical UP state activity, we used calcium imaging and targeted whole-cell recordings of activated neurons in thalamocortical slices of mouse somatosensory cortex. Whereas thalamic stimulation during DOWN states generated multineuronal, synchronized UP states, identical stimulation during UP states had no effect on the subthreshold membrane dynamics of the vast majority of cells or on ongoing multineuronal temporal patterns. Both thalamocortical and corticocortical PSPs were significantly reduced and neuronal input resistance was significantly decreased during cortical UP states – mechanistically consistent with UP state insensitivity. Our results demonstrate that cortical dynamics during UP states are insensitive to thalamic inputs

26th Annual Computational Neuroscience Meeting (CNS*2017): Part 3 - Meeting Abstracts - Antwerp, Belgium. 15–20 July 2017

This work was produced as part of the activities of FAPESP Research,\ud Disseminations and Innovation Center for Neuromathematics (grant\ud 2013/07699-0, S. Paulo Research Foundation). NLK is supported by a\ud FAPESP postdoctoral fellowship (grant 2016/03855-5). ACR is partially\ud supported by a CNPq fellowship (grant 306251/2014-0)

Stress-sensitive antidepressant-like effects of ketamine in the mouse forced swim test.

Major depression is a stress-linked disease with significant morbidity and the anesthetic drug ketamine is of growing interest in the treatment of depression, since in responsive individuals a single dose has rapid (within hours) antidepressant effects that can be sustained for over a week in some instances. This combination of fast action and a therapeutic effect that lasts far beyond the drug's half-life points to a unique mechanism of action. In this reverse translational study, we investigate the degree to which ketamine counteracts stress-related depression-like behavioral responses by determining whether it affects unstressed animals similarly to stressed mice. To test this, male C57BL/6J mice were given a single injection of vehicle (0.9% saline; i.p.), 10 mg/kg ketamine, or 30 mg/kg ketamine, and were tested in the forced swim test (FST) 24 hours and 7 days later, as well as in the open field test on the eighth day. Unstressed mice had normal group housing, environmental enrichment, and experimenter pre-handling (5 days), whereas stressed animals were subjected to chronic mild stress (single housing, reduced enrichment and minimal handling), where some mice also had daily two-week unpredictable chronic stress (UCS). We find that ketamine (24 hours post-injection) decreases immobility and increases mobile (swimming) behavior (antidepressant-like effects) in UCS animals but does the opposite in unstressed mice, similar to recent human findings. In summary, these data suggest that chronic psychological stress interacts with ketamine treatment to modulate its effects in the C57BL/6J mouse FST, which reinforces the relevance of this test, and this strain of mice, to human, stress-induced depression