# Dissemination of 3-photon imaging for chronic cellular imaging across species

> **NIH NIH U01** · ALLEN INSTITUTE · 2022 · $1,025,583

## Abstract

ABSTRACT
Activity-sensitive fluorescent indicators and microscopy have proven valuable tools for measuring neuronal
activity, but most forms of cellular microscopy can produce images of neurons only near the brain surface and
generally only after removal of tissues overlying the brain surface, such as bone. Many neurons in neocortex
are out of reach of cellular microscopy. Here we propose to optimize 3-photon (3P) excitation fluorescence
imaging, a form of cellular microscopy, to enable deeper imaging into the brain. Our aims are to acquire
cellular information from deep cells in flies, mice, cats and macaques.
 After optimizing 3P microscopy, we will facilitate the duplication of our microscope and procedures in other
neuroscience laboratories. We will provide open-access, detailed information on our microscope and
experimental procedures in terminology that’s comprehensible to neuroscientists. In addition, we will work with
one or more microscope manufacturers to ensure commercial availability of our microscope design, enabling
many neuroscience laboratories to use 3P excitation to image deep into the brain with less invasive
preparations, repeatedly over many days, weeks or months.
 To achieve our broad aims, we will pursue five specific aims:
 Specific aim 1: Further optimize fast 3P excitation.
 Specific aim 2: Record calcium signals from central motor circuits in intact, behaving Drosophila
melanogaster.
 Specific aim 3: Study sensory information processing in mouse neocortex and spatial memory in mouse
hippocampus.
 Specific aim 4: Study visual information processing in deep layers of cat and macaque cortex.
 Specific aim 5: Share methods and facilitate availability of a commercial fast 3P microscope.

## Key facts

- **NIH application ID:** 10237131
- **Project number:** 5U01NS115585-02
- **Recipient organization:** ALLEN INSTITUTE
- **Principal Investigator:** Jack Waters
- **Activity code:** U01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $1,025,583
- **Award type:** 5
- **Project period:** 2020-09-01 → 2025-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10237131

## Citation

> US National Institutes of Health, RePORTER application 10237131, Dissemination of 3-photon imaging for chronic cellular imaging across species (5U01NS115585-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10237131. Licensed CC0.

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