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Updated: Jun 15, 2025
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Published on: March 8, 2024
iSECRETE: Integrating Microfluidics and DNA Proximity Amplification for Synchronous Single-Cell Activation and IFN-γ
Ri Lu1,2, Yan Shan Ang3, Ka-Wai Cheung1
1Critical Analytics for Manufacturing of Personalised Medicine IRG, Singapore-MIT Alliance for Research and Technology, Singapore, 138602, Singapore.
Insights
Introducing iSECRETE, a rapid, wash-free platform for single-cell cytokine secretion analysis. This integrated system provides fast, accurate detection of immune responses, advancing biological research.
Area of Science:
- Immunology
- Biotechnology
- Cellular Biology
Background:
- Cytokine dysregulation significantly impacts immune modulation and disease progression.
- Current cytokine profiling methods are often slow, labor-intensive, and fail to capture dynamic immune responses.
Purpose of the Study:
- To develop an integrated platform, iSECRETE, for rapid, single-cell cytokine secretion analysis.
- To enable synchronous cell activation and target-responsive protein detection within a short timeframe.
Main Methods:
- Utilized a multifunctional microfluidic system incorporating a DNA proximity assay (DPA).
- Achieved one-pot homogenous cytokine signal amplification for sensitive detection.
- Demonstrated wash-free, target-responsive protein detection at room temperature.
Main Results:
- iSECRETE provides single-cell IFN-γ cytokine secretion analysis within 30 minutes.
- Achieved a limit of detection of approximately 50 secreted molecules per cell.
- Successfully detected single-cell IFN-γ secretion from CAR T cells upon cancer target exposure within 6 hours.
Conclusions:
- iSECRETE is the fastest single-cell sample-to-result cytokine secretion assay to date.
- The platform offers a powerful tool for understanding biological phenotypes, functions, and pathways under in vivo-like conditions.
- Enables robust analysis across various sample types.
Abstract:
Cytokines, crucial in immune modulation, impact disease progression when their secretion is dysregulated. Existing methods for profiling cytokine secretion suffer from time-consuming and labor-intensive processes and often fail to capture the dynamic nature of immune responses. Here, iSECRETE, an integrated platform that enables synchronous cell activation, wash-free, and target-responsive protein detection for single-cell IFN-γ cytokine secretion analysis within 30 min at room temperature is presented. By incorporating a DNA proximity assay (DPA) into a multifunctional microfluidic system, one-pot homogenous cytokine signal amplification, with a limit of detection of ≈50 secreted molecules per cell is achieved. iSECRETE can robustly handle various sample types that are shown. Two distinct immune activation assay modalities are demonstrated on iSECRETE. Finally, the detection of single-cell IFN-γ secretion as an activation hallmark of chimeric antigen receptor T cells within 6 h of exposure to cancer targets is shown. iSECRETE represents the fastest single-cell sample-to-result cytokine secretion assay to date, providing a powerful tool for advancing the understanding of biological phenotypes, functions, and pathways under in vivo-like conditions.
