# A versatile platform to isolate cells from three-dimensional samples after spatial and temporal profiling

> **NIH NIH R41** · IMAGEN BIOWORKS INC. · 2024 · $333,048

## Abstract

PROJECT SUMMARY/ABSTRACT
Isolating individual cells from mixed populations offers a powerful way to study molecular underpinnings of diseases and
unravel new targets for drug development. Single-cell isolation is also important for directed evolution screens for
engineering biological systems. Despite the growing interest in single-cell isolation techniques from both industry and
academia, current methods have a critical limitation. They fail to isolate individual interesting target cells from tissue or 3D
cultures. Addressing this problem would enable research in a more physiological environment, providing a more accurate
assessment of gene, protein, or cell behaviors and functions. We have recently reported SPOTlight, a versatile all-optical
method for isolating single cells within live cultures based on spatial and temporal properties. In SPOTlight, we optically
tag target cells exhibiting interesting phenotypes through fluorescence photoactivation using a specialized light-patterning
device connected to a microscope. These photoactivated cells, distinguishable by their bright red or green fluorescence, are
then isolated with a FACS machine. However, the current SPOTlight setup relies on a 1-photon widefield microscopy setup,
limiting its utility for 3D samples. The overall objective of this project is to create a 2-photon (2P) microscopy version of
SPOTlight, dubbed 2P-SPOTlight. Our commercialization plan is to provide 2P-SPOTlight as a service to our customers in
industry and academia. In Phase I, we will develop the necessary reagents to allow optical labeling of cells within 3D
cultures using 2-photon light. We will also develop a 2P-SPOTlight assay to isolate spatially distinct subpopulations of
tumor-infiltrating T-cells. We believe this project will satisfy the unmet need to isolate cells from 3D cultures to conduct
downstream spatiotemporal omics.

## Key facts

- **NIH application ID:** 10931998
- **Project number:** 1R41GM154612-01
- **Recipient organization:** IMAGEN BIOWORKS INC.
- **Principal Investigator:** Jihwan Lee
- **Activity code:** R41 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $333,048
- **Award type:** 1
- **Project period:** 2024-09-12 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10931998

## Citation

> US National Institutes of Health, RePORTER application 10931998, A versatile platform to isolate cells from three-dimensional samples after spatial and temporal profiling (1R41GM154612-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10931998. Licensed CC0.

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