Deep Tissue Imaging

Deep tissue imaging comprises methods that visualize cellular structures and activity beneath the surface of intact tissues, enabling neuroscientists to study brain organization in its native context. In optical approaches such as two-photon microscopy, near-infrared light penetrates scattering tissue and produces fluorescence primarily at the focal point, allowing targeted imaging while reducing out-of-focus signal. These techniques can track neurons, glial cells, blood vessels, and neural activity across cortical layers and, with suitable probes and preparations, over time. Deep tissue imaging supports research on circuit function, neurovascular interactions, development, and neurological disease while advancing minimally invasive approaches for observing living brains.

Deep Tissue Imaging - Related Videos

Research

JoVE EoE - Neuroimaging

Deep-Tissue Imaging of a Zebrafish Brain Using a Three-Photon Fluorescence Microscope

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2025

Source: Hontani, Y., et al. Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain. J. Vis. Exp. (2022)This video demonstrates the setup and execution of three-photon imaging in an anesthetized zebrafish brain. The protocol enables deep-tissue visualization of fluorescently labeled neurons, revealing real-time neuronal activity and structures for neuroscience research applications.

In vivo Imaging of Deep Cortical Layers using a Microprism

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Cited by 11 •

2009

Right-angle microprisms inserted into the mouse neocortex allows for deep imaging of multiple cortical layers with a viewpoint typically found in slice. One-millimeter microprisms offer a wide field-of-view (~900 μm) and spatial resolutions sufficient to resolve dendritic spines. We demonstrate layer V neuronal imaging and neocortical vascular imaging using microprisms.

Fluorescent Labeling of Glioma Cells: A Lentiviral Vector-based Transfection Method to Obtain Glioma Cells Expressing Fluorescent Protein for Deep Tissue Imaging

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2025

In this video, we describe a protocol for genetically engineering glioma cells to express infrared fluorescent protein or iRFP. These fluorescent-tagged glioma cells hold utility in deep tissue imaging to understand the development of brain tumors.

Universal Hand-held Three-dimensional Optoacoustic Imaging Probe for Deep Tissue Human Angiography and Functional Preclinical Studies in Real Time

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Cited by 20 •

2014

We provide herein a detailed description of the experimental protocol for imaging with a newly developed hand-held optoacoustic (photoacoustic) system for three-dimensional functional and molecular imaging in real time. The demonstrated powerful performance and versatility may define new application areas of the optoacoustic technology in preclinical research and clinical practice.

Deep Brain Stimulation with Simultaneous fMRI in Rodents

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Cited by 21 •

2014

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

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