# Advanced Single-Cell Protein Analysis with Multiplex in Situ Tagging Array Technology

> **NIH NIH R35** · STATE UNIVERSITY NEW YORK STONY BROOK · 2024 · $393,946

## Abstract

SUMMARY: MIRA
Title: Advanced Single-Cell Protein Analysis with Multiplex in Situ Tagging Array Technology
Single-cell analysis has been the essential approach in understanding cellular machinery, organism development,
and disease mechanisms. Proteins are naturally one of the main targets in single-cell analysis since proteins are
responsible for nearly every task for the cellular life. Unfortunately, proteins are not amplifiable like DNA, and
thus the minuscule amount of proteins on single cells poses a great challenge to the detection techniques. To
date, there is still no single-cell proteome technology available yet that has sufficient sensitivity and scale. We
have innovated a few single-cell protein detection techniques on an ultrahigh-density multiplex in situ tagging
(MIST) array which demonstrates its power for not only analyzing the full spectrum of function proteome in animal
and human cells, but also rapidly detecting cluster of surface proteins on smaller microbial pathogens. The MIST
array is a large-scale monolayer of small-size microbeads with a density much higher than most genome chips,
and the microbeads carry various probes for protein detection. The single-cell MIST (scMIST) technology has
exhibited it ability to quantify in single cells up to 465 functional proteins, which is the highest multiplexity assay
and the most comprehensive mapping of T cell protein markers. The smaller scale of the assay proved its utility
in prediction of sepsis outcomes by analyzing large quantity of primary samples. In the proposed research, we
will extend scMIST to spatially analyze solid tissue specimens by quantifying ~500 functional proteins to
thoroughly assess cellular features, physiological status and functions within a tissue microenvironment. In
addition, we will optimize the scMIST and combine it with artificial intelligence algorithms to precisely prognose
disease occurrence and progression. Last, an explosion-like biomolecular chain reaction will be developed on
the MIST array to rapidly detect few protein copies on single bacteria or viral particles by specifically amplifying
signals for millions to billions of times, with the goal of visually detecting single pathogens within a minute. The
implementation of these advanced single-cell protein detection technologies will catalyze the revolution of
biomedical sciences and fundamentally enhance the precision of disease prognosis. Visual detection of single
pathogens within a minute will bring enormous opportunities in broadly many areas concerned with pathogens.

## Key facts

- **NIH application ID:** 10763760
- **Project number:** 1R35GM151972-01
- **Recipient organization:** STATE UNIVERSITY NEW YORK STONY BROOK
- **Principal Investigator:** Jun Wang
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $393,946
- **Award type:** 1
- **Project period:** 2024-02-01 → 2029-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10763760

## Citation

> US National Institutes of Health, RePORTER application 10763760, Advanced Single-Cell Protein Analysis with Multiplex in Situ Tagging Array Technology (1R35GM151972-01). Retrieved via AI Analytics 2026-05-29 from https://api.ai-analytics.org/grant/nih/10763760. Licensed CC0.

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