by Light Publishing Center, Changchun Institute of Optics, Fine Mechanics And Physics, CAS
Illustration of multi-focus scanning with DEEPscope, imaging a neuron deep within the brain. Credit: by Aaron T. Mok, Tianyu Wang, Shitong Zhao, Kristine E. Kolkman, Danni Wu, Dimitre G Ouzounov, Changwoo Seo, Chunyan Wu, Joseph R. Fetcho, Chris Xu
Researchers at Cornell have unveiled an advanced imaging technology capable of unprecedented deep and wide-field visualization of brain activity at single-cell resolution. The innovative microscope, named DEEPscope, combines two-photon and three-photon microscopy techniques to capture large-scale neural activity and structural details that were previously inaccessible.
Traditional multiphoton microscopy, a cornerstone for deep-tissue imaging, faces significant limitations in imaging depth and field of view, especially in highly scattering biological tissues like the brain. To prevent thermal damage, the imaging depth is usually increased at the expense of an exponentially shrinking field of view, making it challenging to observe large-scale neuronal networks. DEEPscope addresses these constraints by integrating a suite of novel techniques, allowing researchers to visualize extensive brain regions at unprecedented depths.
Key to this advancement is DEEPscope’s adaptive excitation system and multi-focus polygon scanning scheme, which enable efficient fluorescence generation for large field-of-view imaging. These innovations make it possible to achieve high-resolution imaging across a 3.23 x 3.23-mm2 field with an imaging speed that is sufficient for capturing neuronal activity in the deepest cortical layers of mouse brains.
The ability to perform simultaneous two-photon and three-photon imaging further enhances the system’s versatility, allowing detailed exploration of both shallow and deep regions.
(Left) 3D rendering of GCaMP6s-expressing neurons (green) and third harmonic generation signal (magenta) captured by the 3P DEEPscope from 100 to 1,048 µm below the brain surface, showing the cortex, external capsule, and hippocampus. (Middle) YZ maximum intensity projection of the mouse brain. (Right) Digitally zoomed-in images at CA1, 872 µm below the brain surface, and the deepest cortical layer L6, 600 µm below the brain surface. Credit: by Aaron T. Mok, Tianyu Wang, Shitong Zhao, Kristine E. Kolkman, Danni Wu, Dimitre G Ouzounov, Changwoo Seo, Chunyan Wu, Joseph R. Fetcho, Chris Xu
In their study published in the journal eLightthe researchers demonstrated DEEPscope’s capability to image entire cortical columns and subcortical structures with single-cell resolution. They successfully recorded neuronal activities in deep brain regions of transgenic mice, observing over 4,500 neurons in both shallow and deep cortical layers.
Furthermore, DEEPscope enabled whole-brain imaging of adult zebrafish, capturing structural details at depths greater than 1 mm and across a field wider than 3 mm—a first in the field of neuroscience.
“DEEPscope represents a significant advancement in brain imaging technology,” said Aaron Mok, the study’s lead author. “For the first time, we can visualize complex neural circuits in living animals at such a large scale and depth, providing insights into brain function and potentially opening new avenues for neurological research.”
The demonstrated techniques can be readily integrated into existing multiphoton microscopes, making them accessible for widespread use in neuroscience and other fields requiring deep-tissue imaging. By overcoming previous limitations, DEEPscope sets a new standard for large-field, high-resolution, deep imaging of living tissues, promising to advance our understanding of the brain’s intricate networks and their role in health and disease.
More information:
Aaron T. Mok et al, A large field-of-view, single-cell-resolution two- and three-photon microscope for deep and wide imaging, eLight (2024). DOI: 10.1186/s43593-024-00076-4
Provided by Light Publishing Center, Changchun Institute of Optics, Fine Mechanics And Physics, CAS
Citation: New microscope enables deep and wide neuroimaging (2024, November 6) retrieved 7 November 2024 from
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