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1.
Adv Sci (Weinh) ; 9(2): e2103564, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34796701

RESUMO

Cell-type-specific, activity-dependent electrophysiology can allow in-depth analysis of functional connectivity inside complex neural circuits composed of various cell types. To date, optics-based fluorescence recording devices enable monitoring cell-type-specific activities. However, the monitoring is typically limited to a single brain region, and the temporal resolution is significantly low. Herein, a multimodal multi-shank fluorescence neural probe that allows cell-type-specific electrophysiology from multiple deep-brain regions at a high spatiotemporal resolution is presented. A photodiode and an electrode-array pair are monolithically integrated on each tip of a minimal-form-factor silicon device. Both fluorescence and electrical signals are successfully measured simultaneously in GCaMP6f expressing mice, and the cell type from sorted neural spikes is identified. The probe's capability of combined electro-optical recordings for cell-type-specific electrophysiology at multiple brain regions within a neural circuit is demonstrated. The new experimental paradigm to enable the precise investigation of functional connectivity inside and across complex neural circuits composed of various cell types is expected.


Assuntos
Encéfalo/fisiologia , Fenômenos Eletrofisiológicos/fisiologia , Eletrofisiologia/instrumentação , Eletrofisiologia/métodos , Corantes Fluorescentes , Animais , Desenho de Equipamento , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Dispositivos Ópticos
2.
Sci Rep ; 8(1): 15199, 2018 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-30315245

RESUMO

A bright, red-shifted Genetically Encoded Voltage Indicator (GEVI) was developed using a modified version of the fluorescent protein, tdTomato. Dimerization of the fluorescent domain for ArcLight-type GEVIs has been shown to affect the signal size of the voltage-dependent optical signal. For red-shifted GEVI development, tdTomato was split fusing a single dTomato chromophore to the voltage sensing domain. Optimization of the amino acid length and charge composition of the linker region between the voltage sensing domain and the fluorescent protein resulted in a probe that is an order of magnitude brighter than FlicR1 at a resting potential of -70 mV and exhibits a ten-fold larger change in fluorescence (ΔF) upon 100 mV depolarization of the plasma membrane in HEK 293 cells. Unlike ArcLight, the introduction of charged residues to the exterior of dTomato did not substantially improve the dynamic range of the optical signal. As a result, this new GEVI, Ilmol, yields a 3-fold improvement in the signal-to-noise ratio compared to FlicR1 despite a smaller fractional change in fluorescence of 4% per 100 mV depolarization of the plasma membrane. Ilmol expresses well in neurons resolving action potentials in neuronal cultures and reporting population signals in mouse hippocampal acute brain slice recordings. Ilmol is the brightest red-shifted GEVI to date enabling imaging with 160-fold less light than Archon1 for primary neuron recordings (50 mW/cm2 versus 8 W/cm2) and 600-fold less light than QuasAr2 for mouse brain slice recordings (500 mW/cm2 versus 300 W/cm2). This new GEVI uses a distinct mechanism from other approaches, opening an alternate engineering path to improve sensitivity and speed.


Assuntos
Encéfalo/metabolismo , Proteínas Luminescentes/metabolismo , Multimerização Proteica , Sequência de Aminoácidos , Animais , Genes Reporter , Células HEK293 , Hipocampo/metabolismo , Humanos , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Masculino , Camundongos , Modelos Moleculares , Mutação , Neurônios/metabolismo , Imagem Óptica , Conformação Proteica
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