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Updated: Dec 31, 2025
Author Spotlight: Unveiling the Polyfunctionality and Heterogeneity in Immune Responses
Published on: March 8, 2024
Integrated Microfluidic Device for Functional Secretory Immunophenotyping of Immune Cells
Roberto Rodriguez-Moncayo1, Rocio Jimena Jimenez-Valdes1, Alan Mauricio Gonzalez-Suarez1
1Unidad Monterrey , Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional , Parque PIIT , Apodaca , Nuevo León 66628 , Mexico.
Insights
This study introduces a microfluidic platform for automated immune cell analysis. The device measures cytokine secretion, offering potential for disease diagnosis and therapy monitoring.
Area of Science:
- Biomedical Engineering
- Immunology
- Microfluidics
Background:
- Accurate assessment of immune cell function is crucial for diagnosing and monitoring various diseases.
- Current methods for analyzing immune cell secretory function can be complex and lack high throughput.
- Integrated platforms are needed for automated, real-time evaluation of cellular immune responses.
Purpose of the Study:
- To develop and validate a microfluidic platform for the automated assessment of immune cell cytokine secretion.
- To enable simultaneous analysis of multiple immune cell types and responses.
- To improve the efficiency and accuracy of functional immunophenotyping.
Main Methods:
- A microfluidic device with 32 culture chambers, each containing 492 microwells, was designed for immune cell capture.
- A fluorescence sandwich immunoassay utilizing mechanically induced trapping of molecular interactions was employed for cytokine detection.
- Epoxy-modified glass substrates and active mixing were used to enhance biosensing performance.
- Immune cells (monocytes, neutrophils) were captured, cultured, stimulated, and their cytokine secretion (IL-8, TNF-α) analyzed.
Main Results:
- The platform achieved high cell capture efficiency (∼70%) and uniformity (∼90%) for various immune cells.
- Cell viability remained high (up to 96%) for 48 hours in the culture chambers.
- Enhanced biosensing performance was observed with epoxy-modified substrates and active mixing.
- Successful functional secretory analysis of interleukin-8 and tumor necrosis factor alpha was demonstrated.
Conclusions:
- The developed microfluidic platform enables consistent and uniform measurement of cytokine secretion from immune cells.
- This technology holds significant potential for the diagnosis and monitoring of pathologies characterized by altered cytokine profiles.
- The platform offers a robust tool for high-throughput functional immunophenotyping.
Abstract:
Integrated platforms for automatic assessment of cellular functional secretory immunophenotyping could have a widespread use in the diagnosis, real-time monitoring, and therapy evaluation of several pathologies. We present a microfluidic platform with integrated biosensors and culture chambers to measure cytokine secretion from a consistent and uniform number of immune cells. The biosensor relies on a fluorescence sandwich immunoassay enabled by the mechanically induced trapping of molecular interactions method. The platform contains 32 cell culture chambers, each patterned with an array of 492 microwells, to capture and analyze both adherent and nonadherent immune cells. Multiple stimuli can be delivered to a set of culture chambers. Per chamber, we were able to capture consistently 1113 ± 191 of blood-derived monocytes and neutrophils and 348 ± 37 THP-1 monocytes. Good occupancy efficiencies of ∼70% with a uniformity of ∼90% across all of the culture chambers of the device were achieved. Furthermore, we demonstrate that up to 96% of cells remain viable for the first 48 h. The employment of epoxy-modified glass substrates and active mixing enhanced the biosensing performance compared to the use of bare glass and simple diffusion. Finally, we performed functional secretory analysis of interleukin-8 and tumor necrosis factor alpha from human neutrophils and monocytes, stimulated with various doses of lipopolysaccharide and phorbol 12-myristate 13-acetate-ionomycin, respectively. We foresee the employment of our microfluidic platform in the diagnosis of different pathologies where alterations in cytokine secretion patterns can be used as biomarkers.
