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Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for in vivo wide-field imaging of the mouse brain.

ABSTRACT: Large-scale optical measurements have revealed the anatomical and functional connectivity among brain regions underlying brain functions. Here, we describe how to construct a cranial window utilizing a polyethylene-oxide-coated CYTOP (PEO-CYTOP) nanosheet that suppresses bleeding on the brain surface of mice. We demonstrate in vivo two-photon imaging through the PEO-CYTOP nanosheet at the subcellular resolution in the parietal region of the mouse brain. This protocol improves the surgical procedure and expands the optically observable regions, thereby promoting understanding of brain function. For complete details on the use and execution of this protocol, please refer to Takahashi et al. (2020).

SUBMITTER: Takahashi T

PROVIDER: S-EPMC8134076 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for <i>in vivo</i> wide-field imaging of the mouse brain.

Takahashi Taiga T Zhang Hong H Otomo Kohei K Okamura Yosuke Y Nemoto Tomomi T

STAR protocols 20210512 2

Large-scale optical measurements have revealed the anatomical and functional connectivity among brain regions underlying brain functions. Here, we describe how to construct a cranial window utilizing a polyethylene-oxide-coated CYTOP (PEO-CYTOP) nanosheet that suppresses bleeding on the brain surface of mice. We demonstrate <i>in vivo</i> two-photon imaging through the PEO-CYTOP nanosheet at the subcellular resolution in the parietal region of the mouse brain. This protocol improves the surgical ...[more]

PMID: 34027495

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Project description:Significance: Electrophysiological recording and optical imaging are two prevalent neurotechnologies with complementary strengths, the combined application of which can significantly improve our capacity in deciphering neural circuits. Flexible electrode arrays can support longitudinal optical imaging in the same brain region, but their mechanical flexibility makes surgical preparation challenging. Here, we provide a step-by-step protocol by which an ultraflexible nanoelectronic thread is co-implanted with a cranial window in a single surgery to enable chronic, dual-modal measurements. Aim: The method uses 1-μm -thick polymer neural electrodes which conform to the site of implantation. The mechanical flexibility of the probe allows bending without breaking and enables long-lasting electrophysiological recordings of single-unit activities and concurrent, high-resolution optical imaging through the cranial window. Approach: The protocol describes methods and procedures to co-implant an ultraflexible electrode array and a glass cranial window in the mouse neocortex. The implantation strategy includes temporary attachment of flexible electrodes to a retractable tungsten-microwire insertion shuttle, craniotomy, stereotaxic insertion of the electrode array, skull fixation of the cranial window and electrode, and installation of a head plate. Results: The resultant implant allows simultaneous interrogation of brain activity both electrophysiologically and optically for several months. Importantly, a variety of optical imaging modalities, including wide-field fluorescent imaging, two-photon microscopy, and functional optical imaging, can be readily applied to the specific brain region where ultraflexible electrodes record from. Conclusions: The protocol describes a method for co-implantation of ultraflexible neural electrodes and a cranial window for chronic, multimodal measurements of brain activity in mice. Device preparation and surgical implantation are described in detail to guide the adaptation of these methods for other flexible neural implants and cranial windows.

Dataset Information

Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for in vivo wide-field imaging of the mouse brain.

Publications

Protocol for constructing an extensive cranial window utilizing a PEO-CYTOP nanosheet for <i>in vivo</i> wide-field imaging of the mouse brain.

Similar Datasets

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