Animal behavior originates from neuronal activity distributed across brain-wide networks. However, techniques available to assess large-scale neural dynamics in behaving animals remain limited. Here we present compact, chronically implantable, high-density arrays of optical fibers that enable multifiber photometry and optogenetic perturbations across many regions in the mammalian brain. In mice engaged in a texture discrimination task, we achieved simultaneous photometric calcium recordings from networks of 12-48 brain regions, including striatal, thalamic, hippocampal and cortical areas. Furthermore, we optically perturbed subsets of regions in VGAT-ChR2 mice by targeting specific fiber channels with a spatial light modulator. Perturbation of ventral thalamic nuclei caused distributed network modulation and behavioral deficits. Finally, we demonstrate multi-fiber photometry in freely moving animals, including simultaneous recordings from two mice during social interaction. High-density multi-fiber arrays are versatile tools for the investigation of large-scale brain dynamics during behavior.
Despite the growing popularity of blood oxygen level-dependent (BOLD) functional MRI (fMRI), understanding of its underlying principles is still limited. This protocol describes a technique for simultaneous measurement of neural activity using fluorescent calcium indicators together with the corresponding hemodynamic BOLD fMRI response in the mouse brain. Our early work using small-molecule fluorophores in rats gave encouraging results but was limited to acute measurements using synthetic dyes. Our latest procedure combines fMRI with optical detection of cell-type-specific virally delivered GCaMP6, a genetically encoded calcium indicator (GECI). GCaMP6 fluorescence, which increases upon calcium binding, is collected by a chronically implanted optical fiber, allowing longitudinal studies in mice. The chronic implant, placed horizontally on the skull, has an angulated tip that reflects light into the brain and is connected via fiber optics to a remote optical setup. The technique allows access to the neocortex and does not require adaptations of commercial MRI hardware. The hybrid approach permits fiber-optic calcium recordings with simultaneous artifact-free BOLD fMRI with full brain coverage and 1-s temporal resolution using standard gradient-echo echo-planar imaging (GE-EPI) sequences. The method provides robust, cell-type-specific readouts to link neural activity to BOLD signals, as emonstrated for task-free ('resting-state') conditions and in response to hind-paw stimulation. These results highlight the power of fiber photometry combined with fMRI, which we aim to further advance in this protocol. The approach can be easily adapted to study other molecular processes using suitable fluorescent indicators.
In neocortex, each sensory modality engages distinct primary and secondary areas that route information further to association areas. Where signal flow may converge for maintaining information in short-term memory and how behavior may influence signal routing remain open questions. Using wide-field calcium imaging, we compared cortexwide neuronal activity in layer 2/3 for mice trained in auditory and whisker-based tactile discrimination tasks with delayed response. In both tasks, mice were either active or passive during stimulus presentation, engaging in body movements or sitting quietly. Irrespective of behavioral strategy, auditory and tactile stimulation activated spatially segregated subdivisions of posterior parietal cortex (areas A and RL, respectively). In the subsequent delay period, in contrast, behavioral strategy rather than sensory modality determined where short-term memory was located: frontomedially in active trials and posterolaterally in passive trials. Our results suggest behavior-dependent routing of sensory-driven cortical information flow from modality-specific PPC subdivisions to higher association areas..
In this work we present a deperturbation study of the $d\,^3\Pi _g, v=6$d3Πg,v=6 state of C$_{2}$2 by double-resonant four-wave mixing spectroscopy. Accurate line positions of perturbed transitions are unambiguously assigned by intermediate level labeling. In addition, extra lines are accessible by taking advantage of the sensitivity and high dynamic range of the technique. These weak spectral features originate from nearby-lying dark states that gain transition strength through the perturbation process. The deperturbation analysis of the complex spectral region in the (6,5) and (6,4) bands of the Swan system ($d^3\Pi _g-a\,^3\Pi _u$d3Πg−a3Πu) unveils the presence of the energetically lowest high-spin state of C$_{2}$2 in the vicinity of the $d\,^3\Pi _g, v=6$d3Πg,v=6 state. The term energy curves of the three spin components of the d state cross the five terms of the $1\;^5\Pi _g$15Πg state at rotational quantum numbers N ⩽ 11. The spectral complexity for transitions to the v = 6 level of $d\,^3\Pi _g$d3Πg state is further enhanced by an additional perturbation at N = 19 and 21 owing to the $b\, ^3\Sigma _g^-, v=19$b3Σg−,v=19 state. The spectroscopic characterization of both dark states is accessible by the measurement of 122 “window” levels. A global fit of the positions to a conventional Hamiltonian for a linear diatomic molecule yields accurate molecular constants for the quintet and triplet perturber states for the first time. In addition, parameters for the spin-orbit and L-uncoupling interaction between the electronic levels are determined. The detailed deperturbation study unravels major issues of the so-called high-pressure bands of C$_{2}$2. The anomalous nonthermal emission initially observed by Fowler in 1910 [Mon. Not. R. Astron. Soc. 70, 484 (1910)] and later observed in numerous experimental environments are rationalized by taking into account “gateway” states, i.e., rotational levels of the $d\,^3\Pi _g, v=6$d3Πg,v=6 state that exhibit significant $^5\Pi _g$5Πg character through which all population flows from one electronic state to the other.
An optical low-coherence interferometry technique has been used to simultaneously resolve the mode profile and to measure the intermodal dispersion of guided modes of a few-mode fiber. Measurements are performed using short samples of fiber (about 50 cm). There is no need for a complex mode-conversion technique to reach a high interference visibility. Four LP mode groups of the few-mode fiber are resolved. Experimental results and numerical simulations show that the ellipticity of the fiber core leads to a distinct splitting of the degenerate high-order modes in group index. For the first time, to the best of our knowledge, it has been demonstrated that degenerate LP11 modes are much more sensitive to core shape variations than the fundamental modes and that intermodal dispersion of high-
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