Dual fluorophore imaging combined with optical intrinsic signal to acquire neural, metabolic and hemodynamic activity

Wang, Xiaodan; Bice, Annie R.; Padawer-Curry, Jonah A.; Rosenthal, Zachary P.; J, Lee; Bauer, Adam Q.

doi:10.1117/12.2609860

Cited by 3 publications

(9 citation statements)

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“…We perform simultaneous multimodal widefield optical imaging [45] and face videography in awake, head-fixed mice. Fluorescence across the dorsal cortex was measured in 10-minute sessions from a total of N = 7 Thy1-jRGECO1a transgenic mice expressing a red-shifted calcium indicator in excitatory neurons.…”

Section: Resultsmentioning

confidence: 99%

“…3). To do this, we took advantage of a novel multispectral optical imaging platform [46] that simultaneously maps neuronal activity (jRGECO1a fluorescence), oxidative metabolism (as indexed by FAD autofluorescence [67, 68]), and hemodynamics (which approximates the physiological basis for fMRI in humans). We attempted to reconstruct all three biological variables on the basis of delay embedded pupil measurements.…”

Section: Resultsmentioning

confidence: 99%

“…Preprocessing Images were spatially normalized, downsampled (to a resolution of 128x128), co-registered, and affine-transformed to the Paxinos atlas, temporally detrended and spatially smoothed as described previously [91]. Changes in 530nm, and 625nm reflectance were interpreted using the modified Beer-Lambert law to calculate changes in hemoglobin concentration as described previously [45].…”

Section: Datasets and Preprocessingmentioning

confidence: 99%

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Arousal as a universal embedding for spatiotemporal brain dynamics

Raut,

Rosenthal,

Wang

et al. 2023

Preprint

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Neural activity in awake organisms shows widespread and spatiotemporally diverse correlations with behavioral and physiological measurements. We propose that this covariation reflects in part the dynamics of a unified, arousal-related process that regulates brain-wide physiology on the timescale of seconds. This interpretation motivates a surprising prediction: that a single, scalar measurement of arousal (e.g., pupil diameter) should suffice to reconstruct the evolution of multimodal, spatiotemporal measurements of large-scale brain physiology. To test this hypothesis, we perform multimodal, cortex-wide optical imaging and behavioral monitoring in awake mice. We demonstrate that spatiotemporal measurements of neuronal calcium, metabolism, and blood-oxygen can be accurately and parsimoniously modeled from a low-dimensional state-space reconstruction based on the time history of pupil diameter. Extending this framework to behavioral and electrophysiological measurements from the Allen Brain Observatory, we demonstrate the ability to integrate diverse experimental data into a unified generative model via mappings from an intrinsic arousal manifold. Our results support the hypothesis that spontaneous, spatially structured fluctuations in brain-wide physiology — widely interpreted to reflect regionally-specific neural communication — are in large part reflections of an arousal-related process. This enriched view of arousal dynamics has broad implications for interpreting observations of brain, body, and behavior as measured across modalities, contexts, and scales.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Datasets and Preprocessingmentioning

confidence: 99%

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Arousal as a universal embedding for spatiotemporal brain dynamics

Raut,

Rosenthal,

Wang

et al. 2023

Preprint

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“…Here, we describe a new widefield microscope design for mesoscopic imaging of dorsal mouse cortex. Several groups describe widefield calcium imaging with a single fluorescent indicator and concurrent hemodynamic imaging, while other groups describe two-color fluorescence widefield imaging without hemodynamic imaging, while we identified one report describing a system for combined two-color fluorescence and hemodynamic imaging using two cameras (Wang et al 2022). Compared to previous publications, our system has several unique features: First, the system enables quasi-simultaneous imaging of two spectrally distinct fluorophores Second, we use a single detector.…”

Section: Discussionmentioning

confidence: 99%

“…Several groups incorporated a second fluorescence channel with excitation at 560-590 nm to enable two-color fluorescence imaging of red fluorescent calcium indicators like jRCaMP1b or jRGECO1a in combination with either EGFP-derived indicators such as GCaMP6 (Gribizis et al 2019), GRABACh (Lohani et al 2022), or flavoprotein autofluorescence (Wang et al 2022, Raut et al 2023. Widefield imaging systems dedicated to the investigation of neurovascular interactions typically combine imaging of green (Ma et al 2016b, Matsui et al 2016, Wright et al 2017, Mitra et al 2018, Valley et al 2020, Sunil et al 2023, red fluorescent indicators (Park et al 2020, Raut et al 2023, Shahsavarani et al 2023, or green and red fluorescent indicators (Wang et al 2022) with reflectance imaging for D[HbO] and D [HbR] estimation at two or three wavelengths, typically in the range of 525-565 nm and 590-625 nm.…”

Section: Introductionmentioning

confidence: 99%

Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination

Doran,

Fomin-Thunemann,

Tang

et al. 2023

Preprint

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SignificanceWidefield microscopy of the entire dorsal part of mouse cerebral cortex enables large-scale (“mesoscopic”) imaging of neuronal activity with fluorescent indicators as well as hemodynamics via oxy- and deoxyhemoglobin absorption. Versatile and cost-effective imaging systems are needed for large-scale, color-multiplexed imaging of multiple fluorescent and intrinsic contrasts.AimDevelop a system for mesoscopic imaging of two fluorescent and two reflectance channels.ApproachExcitation of red and green fluorescence is achieved through epi-illumination. Hemoglobin absorption imaging is achieved using 525- and 625-nm LEDs positioned around the objective lens. An aluminum hemisphere placed between objective and cranial window provides diffuse illumination of the brain. Signals are recorded sequentially by a single sCMOS detector.ResultsWe demonstrate performance of our imaging system by recording large-scale spontaneous and stimulus-evoked neuronal, cholinergic, and hemodynamic activity in awake head-fixed mice with a curved “crystal skull” window expressing the red calcium indicator jRGECO1a and the green acetylcholine sensor GRABACh3.0. Shielding of illumination light through the aluminum hemisphere enables concurrent recording of pupil diameter changes.ConclusionsOur widefield microscope design with single camera can be used to acquire multiple aspects of brain physiology and is compatible with behavioral readouts of pupil diameter.

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Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination

Doran,

Fomin-Thunemann,

Tang

et al. 2024

Neurophoton.

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Widefield microscopy of the entire dorsal part of mouse cerebral cortex enables large-scale ("mesoscopic") imaging of different aspects of neuronal activity with spectrally compatible fluorescent indicators as well as hemodynamics via oxy-and deoxyhemoglobin absorption. Versatile and cost-effective imaging systems are needed for large-scale, color-multiplexed imaging of multiple fluorescent and intrinsic contrasts. Aim:We aim to develop a system for mesoscopic imaging of two fluorescent and two reflectance channels.Approach: Excitation of red and green fluorescence is achieved through epi-illumination. Hemoglobin absorption imaging is achieved using 525-and 625-nm lightemitting diodes positioned around the objective lens. An aluminum hemisphere placed between objective and cranial window provides diffuse illumination of the brain. Signals are recorded sequentially by a single sCMOS detector. Results:We demonstrate the performance of our imaging system by recording large-scale spontaneous and stimulus-evoked neuronal, cholinergic, and hemodynamic activity in awake, head-fixed mice with a curved "crystal skull" window expressing the red calcium indicator jRGECO1a and the green acetylcholine sensor GRAB ACh3.0 . Shielding of illumination light through the aluminum hemisphere enables concurrent recording of pupil diameter changes.Conclusions: Our widefield microscope design with a single camera can be used to acquire multiple aspects of brain physiology and is compatible with behavioral readouts of pupil diameter.

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Dual fluorophore imaging combined with optical intrinsic signal to acquire neural, metabolic and hemodynamic activity

Cited by 3 publications

References 0 publications

Arousal as a universal embedding for spatiotemporal brain dynamics

Arousal as a universal embedding for spatiotemporal brain dynamics

Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination

Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination

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