Hydrogels are widely used in nerve tissue repair and show good histocompatibility. There remain, however, challenges with hydrogels for applications related to neural signal recording, which requires a tissue-like biomechanical property, high optical transmission, and low impedance. Here, we describe a transparent hydrogel that is highly biocompatible and has a low Young's modulus (0.15 MPa). Additionally, it functions well as an implantable electrode, as it conformably adheres to brain tissue, results in minimal inflammation and has a low impedance of 150 Ω at 1 kHz. Its high transmittance, corresponding to 93.35% at a wavelength of 300 nm to 1100 nm, supports its application in two-photon imaging. Consistent with these properties, this flexible multimodal transparent electrophysiological hydrogel (MTEHy) electrode was able to record neuronal Ca2+ activity using miniature two-photon microscopy. It also used to monitor electrocorticogram (ECoG) activity in real time in freely moving mice. Moreover, its compatibility with magnetic resonance imaging (MRI), indicates that MTEHy is a new tool for studying activity in the cerebral cortex.
Functional network topography of the medial entorhinal cortex
The medial entorhinal cortex (MEC) creates a map of local space, based on the firing patterns of grid, head-direction (HD), border, and object-vector (OV) cells. How these cell types are organized anatomically is debated. In-depth analysis of this question requires collection of precise anatomical and activity data across large populations of neurons during unrestrained behavior, which neither electrophysiological nor previous imaging methods fully afford. Here, we examined the topographic arrangement of spatially modulated neurons in the superficial layers of MEC and adjacent parasubiculum using miniaturized, portable two-photon microscopes, which allow mice to roam freely in open fields. Grid cells exhibited low levels of co-occurrence with OV cells and clustered anatomically, while border, HD, and OV cells tended to intermingle. These data suggest that grid cell networks might be largely distinct from those of border, HD, and OV cells and that grid cells exhibit strong coupling among themselves but weaker links to other cell types.
Itch perception is reflected by neuronal ignition in the primary somatosensory cortex
Multiple cortical areas including the primary somatosensory cortex (S1) are activated during itch signal processing, yet cortical representation of itch perception remains unknown. Using novel miniature two-photon microscopic imaging in free-moving mice, we investigated the coding of itch perception in S1. We found that pharmacological inactivation of S1 abolished itch-induced scratching behavior, and the itch-induced scratching behavior could be well predicted by the activity of a fraction of layer 2/3 pyramidal neurons, suggesting that a subpopulation of S1 pyramidal neurons encoded itch perception, as indicated by immediate subsequent scratching behaviors. With a newly established optogenetics-based paradigm that allows precisely controlled pruritic stimulation, we found that a small fraction of S1 neurons exhibited an ignition-like pattern at the detection threshold of itch perception. Our study revealed the neural mechanism underlying itch perceptual coding in S1, thus paving the way for the study of cortical representation of itch perception at the single-neuron level in freely moving animals.
Miniature two-photon microscopy for enlarged field-of-view, multi-plane and long-term brain imaging
We have developed a miniature two-photon microscope equipped with an axial scanning mechanism and a long-working-distance miniature objective to enable multi-plane imaging over a volume of 420 × 420 × 180 μm3 at a lateral resolution of ~1 μm. Together with the detachable design that permits long-term recurring imaging, our miniature two-photon microscope can help decipher neuronal mechanisms in freely behaving animals.
An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRABACh (GPCR-activation-based ACh) sensor to achieve substantially improved sensitivity in ACh detection, as well as reduced downstream coupling to intracellular pathways. The improved version of the ACh sensor retains the subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for ACh of its predecessor. Using this sensor, we revealed compartmental ACh signals in the olfactory center of transgenic flies in response to external stimuli including odor and body shock. Using fiber photometry recording and two-photon imaging, our ACh sensor also enabled sensitive detection of single-trial ACh dynamics in multiple brain regions in mice performing a variety of behaviors.
Miniature Fluorescence Microscopy for Imaging Brain Activity in Freely-Behaving Animals
An ultimate goal of neuroscience is to decipher the principles underlying neuronal information processing at the molecular, cellular, circuit, and system levels. The advent of miniature fluorescence microscopy has furthered the quest by visualizing brain activities and structural dynamics in animals engaged in self-determined behaviors. In this brief review, we summarize recent advances in miniature fluorescence microscopy for neuroscience, focusing mostly on two mainstream solutions – miniature single-photon microscopy, and miniature two-photon microscopy. We discuss their technical advantages and limitations as well as unmet challenges for future improvement. Examples of preliminary applications are also presented to reflect on a new trend of brain imaging in experimental paradigms involving body movements, long and complex protocols, and even disease progression and aging.
B2,Building 16,Treehouse,73 Tanmi Road,
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P. R. CHINA
