# High-Throughput, Multiplexing-Ready Intracellular Pressure Probes

> **NIH NIH R21** · JOHNS HOPKINS UNIVERSITY · 2022 · $268,758

## Abstract

PROJECT SUMMARY
It is known that at the level of a single cell, intracellular pressure governs motility, shape, volume, and proliferation.
Mounting evidence suggests that pressure can vary within a single cell and the compartmentalization of pressure
may be essential for dynamic cell function. Thus, it is essential to develop approaches to map the heterogeneous
intracellular pressure within a single cell at submicron resolution given the technical limitations of the current
technology. Specifically, it is challenging to study how intracellular pressure regulates cellular processes, such
as protrusion of the cell cortex, heterogeneously and dynamically, to result in certain phenotypes, such as
directional migration. This challenge stems from the lack of nano-sized sensors that are compatible for high-
throughput multiplexing imaging so that local intracellular pressure and other dynamic processes can be
simultaneously measured across the cell.
Herein we propose to develop a high-throughput, multiplexing-ready intracellular probe in the form of nano-
sized liposomes enclosed by DNA-based scaffold with aquaporin molecules distributed in the lipid bilayer.
Joining the DNA scaffold and the aquaporin-embedded liposome are elastic DNA tethers conjugated with
Foster Resonance Energy Transfer (FRET) donor and acceptor fluorophores at prescribed spacing, which
extend or contract as the result of pressure-dependent changes to liposome volume. The nano-sized pressure
sensor, coined “aquaporin-laced liposome pressure sensor (ALPS)”, will be delivered to the cytoplasm in
quantity. Upon pressure changes in the cytoplasm, the internalized ALPS will change its volume by water efflux
or influx through the aquaporin, while the DNA scaffold stabilizes the liposome to prevent collapse or rupture.
As a proof of concept, we will then use ALPS to map the dynamic pressure field induced within single cells
using compartmentalized pressure to migrate within 3D matrix; the results will be compared to the direct
measurements obtained by 0.5-μm micro-electrodes with limited spatial resolution, the current state-of-art. If
successful, we will generate a novel tool for measuring intracellular pressure with unprecedented
spatiotemporal resolution, which promises to provide insights on how local intracellular pressure changes
dynamically as cells navigate the 3D terrain.

## Key facts

- **NIH application ID:** 10431428
- **Project number:** 1R21GM146105-01
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Yun Chen
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $268,758
- **Award type:** 1
- **Project period:** 2022-09-17 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10431428

## Citation

> US National Institutes of Health, RePORTER application 10431428, High-Throughput, Multiplexing-Ready Intracellular Pressure Probes (1R21GM146105-01). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10431428. Licensed CC0.

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