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1.
Nat Methods ; 20(10): 1581-1592, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37723246

RESUMO

Here we report SUPPORT (statistically unbiased prediction utilizing spatiotemporal information in imaging data), a self-supervised learning method for removing Poisson-Gaussian noise in voltage imaging data. SUPPORT is based on the insight that a pixel value in voltage imaging data is highly dependent on its spatiotemporal neighboring pixels, even when its temporally adjacent frames alone do not provide useful information for statistical prediction. Such dependency is captured and used by a convolutional neural network with a spatiotemporal blind spot to accurately denoise voltage imaging data in which the existence of the action potential in a time frame cannot be inferred by the information in other frames. Through simulations and experiments, we show that SUPPORT enables precise denoising of voltage imaging data and other types of microscopy image while preserving the underlying dynamics within the scene.


Assuntos
Microscopia , Redes Neurais de Computação , Razão Sinal-Ruído , Distribuição Normal , Processamento de Imagem Assistida por Computador/métodos
2.
bioRxiv ; 2024 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-39282466

RESUMO

Established methods for imaging the living mammalian brain have, to date, taken optical properties of the tissue as fixed; we here demonstrate that it is possible to modify the optical properties of the brain itself to significantly enhance at-depth imaging while preserving native physiology. Using a small amount of any of several biocompatible materials to raise the refractive index of solutions superfusing the brain prior to imaging, we could increase several-fold the signals from the deepest cells normally visible and, under both one-photon and two-photon imaging, visualize cells previously too dim to see. The enhancement was observed for both anatomical and functional fluorescent reporters across a broad range of emission wavelengths. Importantly, visual tuning properties of cortical neurons in awake mice, and electrophysiological properties of neurons assessed ex vivo, were not altered by this procedure.

3.
J Vis Exp ; (194)2023 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-37184275

RESUMO

As a vertebrate model animal, larval zebrafish are widely used in neuroscience and provide a unique opportunity to monitor whole-brain activity at the cellular resolution. Here, we provide an optimized protocol for performing whole-brain imaging of larval zebrafish using three-dimensional fluorescence microscopy, including sample preparation and immobilization, sample embedding, image acquisition, and visualization after imaging. The current protocol enables in vivo imaging of the structure and neuronal activity of a larval zebrafish brain at a cellular resolution for over 1 h using confocal microscopy and custom-designed fluorescence microscopy. The critical steps in the protocol are also discussed, including sample mounting and positioning, preventing bubble formation and dust in the agarose gel, and avoiding motion in images caused by incomplete solidification of the agarose gel and paralyzation of the fish. The protocol has been validated and confirmed in multiple settings. This protocol can be easily adapted for imaging other organs of a larval zebrafish.


Assuntos
Encéfalo , Imageamento Tridimensional , Microscopia Intravital , Microscopia de Fluorescência , Neuroimagem , Peixe-Zebra , Animais , Encéfalo/diagnóstico por imagem , Imageamento Tridimensional/instrumentação , Imageamento Tridimensional/métodos , Neuroimagem/instrumentação , Neuroimagem/métodos , Sefarose , Microscopia Intravital/instrumentação , Microscopia Intravital/métodos
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