# A 3D super-resolved platform for rapid and highly multiplexed proteomics

> **NIH NIH R44** · PANLUMINATE INC. · 2024 · $1,219,907

## Abstract

Project Summary
Addressing the complexities of brain function and disorders at the cellular and molecular level
requires a spatial omics technology, capable of mapping the 3D distribution of specific
cell markers within tissue at the subcellular scale. While spatial proteomics technologies
have revolutionized in situ molecular profiling of tissues, limitations in resolution, epitope
accessibility, sample degradation, and 3D imaging capability have hindered their widespread
impact. This is the focus of our project, which proposes to combine two groundbreaking
microscopy technologies, pan-Single Step Expansion Microscopy (pan-SSExM) and
FLASH-PAINT, into a revolutionary new method named pan-OptiX S3D. Our optimized
pan-SSExM protocol offers tunable physical sample expansion between 1.5-fold and 8-fold
linearly, while retaining the proteome for subsequent labeling. FLASH-PAINT, on the other hand,
allows rapid and unlimited 3D multiplexed protein labeling without any sample damage. By
synergizing these technologies, pan-OptiX S3D promises subcellular and multicolor optical
imaging, in 3D, across whole tissue sections, with no label limitations due to degradation or
steric hindrance. Panluminate, Inc. is a leading company in the Spatial Biology and Tissue
Expansion fields. Our proposed project aims to develop pan-OptiX S3D and validate it in rodent
brain tissue to enable analysis of neural networks and pathologies in the brain. We specifically
propose to (1) develop novel FLASH probes specifically designed for compatibility with 5-fold
tissue expansion imaging, and (2) automate sample expansion and microfluidics multiplexing
with 3D microscopy and software integration. Ultimately, the developments proposed here will
enable panluminate to disseminate pan-OptiX S3D sample preparation and imaging to every
research and biotech lab around the world.

## Key facts

- **NIH application ID:** 11008498
- **Project number:** 2R44MH128999-02
- **Recipient organization:** PANLUMINATE INC.
- **Principal Investigator:** Ons M'Saad
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $1,219,907
- **Award type:** 2
- **Project period:** 2022-09-16 → 2026-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11008498

## Citation

> US National Institutes of Health, RePORTER application 11008498, A 3D super-resolved platform for rapid and highly multiplexed proteomics (2R44MH128999-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/11008498. Licensed CC0.

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