Deep Tissue Imaging
Deep Tissue Imaging - Related Videos
03:22
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.
09:45
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.
07:29
Deep Vascular Imaging in the Eye with Flow-Enhanced Ultrasound
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Cited by 2 •
2021
We present a non-invasive ultrasound technique for generating three-dimensional angiographies in the eye without the use of contrast agents.
08:26
Deep-Tissue Three-Photon Fluorescence Microscopy in Intact Mouse and Zebrafish Brain
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Cited by 8 •
2022
Three-photon microscopy enables high-contrast fluorescence imaging deep in living biological tissues, such as mouse and zebrafish brains, with high spatiotemporal resolution.
07:44
A Head-Mounted Optically Transparent Skull (HOTS) Window For Deep Transcranial Imaging of the Mouse Cortex
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2026
This protocol describes a head-mounted optically transparent skull (HOTS) window based on an optimized two-step skull-clearing procedure. This approach yields a highly transparent skull, enabling two-photon imaging of the adult mouse cerebral cortex to depths of up to ~800 µm.
FAQs
Near-infrared excitation can travel through scattering tissue, while fluorescence is produced primarily at the focal point. This confines the detected signal to the selected location and reduces out-of-focus fluorescence. In practice, that localization supports targeted observation of structures and activity within distinct cortical layers.
The structures and activity available for observation depend on using suitable probes and preparations for the tissue and imaging goal. These choices determine whether investigators can follow neurons, glial cells, blood vessels, or neural activity beneath the surface. Matching the preparation to the target therefore expands the information obtained.
Suppressing signal from outside the focal region helps separate nearby cellular structures and activity along the imaging path. This is particularly valuable in intact tissue, where scattering can complicate interpretation. The result is more targeted visualization of organization across cortical layers rather than an undifferentiated fluorescent signal.
Investigators align the imaging approach with the structure or activity they want to observe, then select suitable probes and tissue preparations. The optical method can be configured for targeted observation beneath the surface, including cortical layers. This planning determines whether measurements emphasize cells, blood vessels, or neural activity.
Depending on the probe and preparation, imaging can follow neurons, glial cells, blood vessels, and neural activity. Because observations can be made in intact tissue and, in suitable settings, over time, the approach can connect cellular signals with changing circuit or vascular states. This supports studies of brain function in its native context.
It enables investigation of brain organization, circuit function, neurovascular interactions, development, and neurological disease. Imaging neurons alongside glial cells and blood vessels can provide context for how cellular activity relates to vascular or circuit processes. Its ability to observe living brains over time also supports questions about changing neural systems.