# Nanoplasmonic Spatiotemporal Imaging of Single-Cell Protein Secretion and Intercellular Communication

> **NIH NIH R21** · NEW YORK UNIVERSITY · 2024 · $196,819

## Abstract

ABSTRACT
The ability to probe the temporal profile of the protein secretion behavior of individual immune cells will impact
future immunology, cell biology, and even infectious disease diagnosis. Knowledge of the ordering and timing of
cytokines (water-soluble proteins essential for intercellular signaling) secreted by activated T cells can
additionally provide the means to discriminate subsets of differentiated T cells by function. Here, the temporal
information is one of the pieces of the whole puzzle in monitoring the behavior of the immune system. The other
critical piece is the cytokine-mediated interplay between different cell types, which involves spatial transport of
cytokines between cells. Putting both pieces of the puzzle together allows us to capture the full picture of the
cytokine release dynamics and cytokine-mediated interactions of cells, which allows us to fully understand the
intercellular signaling processes underlying immunity. However, no study has yet obtained such a picture due to
the lack of a technology for real-time sensing of intercellular cytokine-mediated signaling processes at high
spatial resolution. This research aims to develop a novel label-free imaging technique to fully understand cellular
behaviors during cytokine-mediated activation and communication at a single-cell level. Our approach will
employ biosensors consisting of plasmonic nanoantenna structures, each specifically targeting a particular
cytokine species. We will integrate these biosensors in a microfluidic system incorporating an array of
sample/reagent-flow channels and single-cell trapping microwells. The microfluidic sensor integration will provide
the ability to capture, manipulate, and activate single cells for cell-to-cell communications on a single chip and
to obtain the spatiotemporal profile of cellular cytokine secretion processes in real time, both in a massively,
parallel manner. We will also develop a theoretical algorithm that allows us to extract the quantitative values of
the local cytokine concentration distributions from measured image intensities. SA 1: We will create highly
ordered, high-density plasmonic nanoantenna biosensor arrays, each functionalized by highly selective
aptamers against targeted cytokines. SA 2: We will integrate the aptamer-conjugated plasmonic nanoantenna
arrays into a single-cell manipulation microfluidic system and achieve real-time single-cell secretion imaging at
high throughput. SA 3: We will develop a two-mode (fluorescence and dark-field) microscopy imaging technique
to image spatiotemporal cytokine secretomic profile patterns and cell surface sytokine binding sites. Using this
technique, we will study the IL-6-mediated dynamic intercellular communication between individual human
hepatoma Hep3b cells and CD 4+ T cells.

## Key facts

- **NIH application ID:** 10935968
- **Project number:** 5R21GM151528-02
- **Recipient organization:** NEW YORK UNIVERSITY
- **Principal Investigator:** Katsuo Kurabayashi
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $196,819
- **Award type:** 5
- **Project period:** 2023-09-30 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10935968

## Citation

> US National Institutes of Health, RePORTER application 10935968, Nanoplasmonic Spatiotemporal Imaging of Single-Cell Protein Secretion and Intercellular Communication (5R21GM151528-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10935968. Licensed CC0.

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