# Three-Photon Microscope for High-Resolution Deep Tissue Imaging

> **NIH NIH S10** · MASSACHUSETTS GENERAL HOSPITAL · 2021 · $599,899

## Abstract

Project Summary
Three-photon microscopy (3PM) is an optical imaging technology that enables significantly deeper imaging in a
scattering biological tissue than the current leading standard two-photon microscopy (2PM), while providing
additional contrast mechanisms and enhancing spatial resolution. This technology is made possible by advances
in long-wavelength, tunable, high-pulse-energy femtosecond lasers and corresponding modifications in 2PM
setups to accommodate a novel excitation regime.
As a direct result of these advancements, 3PM imaging depth in rodent brain is greatly extended with respect to
2PM imaging depth—from ~700 μm to more than 1,200 μm. Due to such a large improvement, we anticipate
that 3PM will allow breakthrough studies of the structure (e.g., vascular, glial, and neural architecture) and
function (e.g., oxygenation, calcium signaling, and cellular interactions) in preclinical in vivo studies, with cellular-
size resolution in previously inaccessible tissue layers, such as the subcortical white matter and the
hippocampus. The 11 NIH-funded projects listed in this proposal will significantly benefit from these imaging
capabilities. These projects span a diverse range of research topics relevant to NIMH mission, including
improvement of the quantitativeness of the clinical imaging modalities and better understanding of the
mechanisms involved in development of various brain conditions.
The Martinos Center for Biomedical Imaging and Massachusetts General Hospital constitute a diverse
community of researchers, many of whom are funded with NIH support, performing cutting edge biomedical
research that will further advance our fundamental understanding of human health and disease. This proposed
3PM imaging system will represent the first instrument of its kind in the Boston area (and probably Northeast
US) that is available within an Imaging Core. This cutting-edge instrumentation will allow our research community
to significantly enhance various research studies relevant to NIMH mission, and it will also allow the Martinos
Center to continue prospering as a leader for advanced biomedical imaging.

## Key facts

- **NIH application ID:** 10282619
- **Project number:** 1S10MH124730-01A1
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** Sava Sakadzic
- **Activity code:** S10 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $599,899
- **Award type:** 1
- **Project period:** 2021-08-15 → 2022-08-14

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10282619

## Citation

> US National Institutes of Health, RePORTER application 10282619, Three-Photon Microscope for High-Resolution Deep Tissue Imaging (1S10MH124730-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10282619. Licensed CC0.

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