# A multi-plane 3-photon microscope for volume imaging in NHP cortex

> **NIH NIH R34** · ALLEN INSTITUTE · 2021 · $801,704

## Abstract

PROJECT ABSTRACT
Recent advances in microscopy permit volume imaging - the near-simultaneous imaging of many neurons in a circuit -
with 2-photon (2P) excitation, yielding new insights into the circuits underlying relatively simple behaviors. Volume
imaging techniques and 2P excitation have been limited mainly to small animals such as rodents, including studies of
mouse cortex. Mouse cortex differs from neocortex in non-human primates and humans and it is unlikely that studies of
mouse cortex will be sufficient to understand the role of cortex in humans.
 In this R34 application, we propose to build a multi-plane 3-photon (3P) fluorescence microscope to perform volume
imaging in macaques. Unlike 2P excitation, 3P excitation supports imaging deep into macaque cortex. In a future R01
application, we will use this 3P microscope to image large volumes of macaque primary visual cortex, studying how
motion information is pooled across neurons in layer 4B and pairing 3P microscopy with serial-section electron
microscopy to study the processing of color information, particularly in layer 4Cβ.
 We will pursue four specific aims:
 - Specific aim 1: Construct and test a multi-plane 3P microscope
 - Specific aim 2: Measure visual tuning of ~50,000 neurons in layers 1-4 in a macaque.
 - Specific aim 3: Extend multi-plane 3P imaging through layers 5 and 6 of macaque cortex.
 - Specific aim 4: Make hardware designs, software and protocols freely available.

## Key facts

- **NIH application ID:** 10205806
- **Project number:** 1R34NS121766-01
- **Recipient organization:** ALLEN INSTITUTE
- **Principal Investigator:** Jack Waters
- **Activity code:** R34 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $801,704
- **Award type:** 1
- **Project period:** 2021-04-15 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10205806

## Citation

> US National Institutes of Health, RePORTER application 10205806, A multi-plane 3-photon microscope for volume imaging in NHP cortex (1R34NS121766-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10205806. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*