Non-invasive cellular-resolution deep imaging of brain structure and function

The National Institute of Neurological Disorders and Stroke awarded Cornell University a $686,023 Cooperative Agreement on July 8, 2026, to develop non-invasive optical imaging technology for visualizing brain structure and function at the cellular level.

Cornell will advance three-photon microscopy (3PM) imaging techniques and develop new lasers to increase three-photon fluorescence signal by approximately 10,000 times. This advancement will enable visualization of mouse brain activity with sub-cellular spatial resolution at 2 millimeters depth—nearly twice the depth achievable with current activity imaging. The work addresses fundamental technological barriers in 3PM that have prevented it from reaching its theoretical potential. Cornell will partner with industry to explore commercialization and broad dissemination of the technology.

The award funds research under the Brain Research through Advancing Innovative Neurotechnologies (BRAIN) Initiative (CFDA 93.372), which supports innovative neurotechnologies to produce dynamic pictures of how individual cells and neural circuits interact in time and space, ultimately enabling new treatments for brain disorders.

The work is performed at Cornell's location in New York and runs through June 30, 2030. Principal investigator Chris Xu and co-investigators Frank Wise and Nilay Yapici lead the research program.

Abstract

Optical imaging is non-invasive and provides the high spatial resolution necessary to resolve individual neurons and neuronal processes. However, acquiring images through significant depths of the brain is no easy task since brain tissue is extremely heterogeneous, resulting in strong scattering by the various tissue components. Multiphoton microscopy (MPM), which was first demonstrated more than three decades ago, has dramatically extended the depth penetration of high-resolution optical imaging. For example, when imaging mouse brain activity with gcamps, 920-NM 2-photon microscopy (2PM) increased the imaging depth to ~ 0.7 MM, and the more recent development of 1300 NM 3-photon microscopy (3PM) further increased the penetration depth to ~ 1.1 MM. While theory and experiments have shown that 920 NM 2PM has reached its fundamental depth limit, the depth limit for 1300 NM 3PM is about twice the deepest imaging demonstrated today. We have identified the major technological barriers that kept existing 3PM from reaching its full potential and discovered new routes to address them. Through the development of lasers and new imaging techniques, this research program will increase the three-photon fluorescence signal by approximately 10,000X; this will ultimately enable visualization of mouse brain activity with sub-cellular spatial resolution at 2 MM depth, which is nearly twice the depth of the deepest activity imaging today. The research involves close interactions between the PI (chris XU) and CO-investigators (frank wise and nilay yapici). Furthermore, we will work with industry partners to explore commercialization of the technology, which will provide a direct path to broad dissemination. The successful completion of this program will dramatically increase the optical imaging depth and enable investigation of dynamic biological phenomena at the single cell level in intact organs and animal models that are completely beyond the reach of any existing imaging techniques.