# Instrumentation platform for 3D pathology with open-top light-sheet microscopy

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 2022 · $469,431

## Abstract

Summary
The clinical gold-standard method for interrogating tissue specimens, slide-based (2D) histopathology, is based
on centuries-old technologies with many inherent limitations. Recent technological advances have demonstrated
the feasibility of achieving high-throughput slide-free 3D histology of biopsy and surgical specimens. In
comparison to conventional slide-based histology, nondestructive 3D histology has the potential to provide a
transformative improvement in diagnostic pathology performance for a number of reasons: (1) vastly greater
(>100X) sampling of tissue specimens, (2) volumetric imaging of 3D cell distributions and tissue structures that
are prognostic and predictive, (3) nondestructive imaging, which allows valuable biopsy specimens to be used
for downstream biomarker assessment, and (4) a simplified process with cost benefits for healthcare institutions
and payers. In recent years, we have developed a technology, open-top light-sheet (OTLS) microscopy, to
enable high-throughput nondestructive 3D histology of ex vivo specimens. Our first generations of OTLS
microscopes and imaging protocols demonstrated the ability to reliably image a variety of optically cleared clinical
tissue specimens (surgical excisions and biopsies) in a nondestructive manner that does not interfere with
conventional pathology methods. Here, we propose to develop a multi-resolution hybrid OTLS microscope (Aim
1), based on a novel non-orthogonal dual-objective (NODO) architecture, which will be superior in every regard
to our previous systems, including resolution (and range of resolutions), imaging depth, and compatibility with
nearly all clearing/labeling protocols and sample-holder materials (insensitivity to refractive-index mismatch).
Furthermore, we will develop innovative pre-imaging methods to automate and standardize the tissue-labeling
and clearing process for a robust fluorescent analog of H&E staining (Aim 2). Finally, we will develop post-
imaging technologies for image-guided macro-dissection of thick tissues, which we will show has the ability to
significantly improve the sensitivity of genomic assays (Aim 3). Collectively, our project aims are designed to
extend current 2D pathology workflows into 3D to minimize clinical-adoption barriers. A rapid translational
pathway exists through a 3D-pathology-services company (Lightspeed Microscopy Inc.) that has licensed our
entire 3D pathology IP portfolio. As part of this larger translational effort, clinical studies are ongoing in our labs,
along with development of AI-analysis methods for clinical decision-support (i.e. prognostication and prediction
of treatment response). The instrumentation platform developed in this project will directly support a number of
future disease-focused clinical studies to demonstrate the value of 3D pathology for the precision treatment of
diverse conditions such as kidney disease, neurodegenerative diseases, and various forms of cancer.

## Key facts

- **NIH application ID:** 10434718
- **Project number:** 5R01EB031002-02
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Jonathan T.C. Liu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $469,431
- **Award type:** 5
- **Project period:** 2021-07-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10434718

## Citation

> US National Institutes of Health, RePORTER application 10434718, Instrumentation platform for 3D pathology with open-top light-sheet microscopy (5R01EB031002-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10434718. Licensed CC0.

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