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Updated: Aug 19, 2025

Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay

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Sequential Reagent Release from a Layered Tablet for Multistep Diagnostic Assays.

Vanessa Y C Li^1,2, Buddhisha Udugama^1,2, Pranav Kadhiresan^1,2

¹Institute of Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S 3G9, Canada.

Analytical Chemistry

|December 1, 2022

Summary

A novel multilayer tablet simplifies complex diagnostic assays by sequentially releasing prequantified reagents. This innovation enables multistep diagnostic testing without user intervention, making assays more accessible.

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Last Updated: Aug 19, 2025

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Area of Science:

Biomaterials Engineering
Analytical Chemistry
Point-of-Care Diagnostics

Background:

Multistep diagnostic assays require precise reagent addition, hindering use by untrained individuals.
Current methods rely on trained technicians and specialized instrumentation for reagent handling.

Purpose of the Study:

To develop a user-friendly system for automated, sequential reagent delivery in complex diagnostic assays.
To simplify multistep diagnostic processes for broader accessibility.

Main Methods:

Designed a multilayer tablet using cellulose-grade polymers (microcrystalline cellulose, hydroxypropyl cellulose).
Controlled reagent release rates by modulating layer compression force and chemical formulation.
Validated the tablet system with two distinct assays: nucleic acid detection and nitrite ion measurement.

Main Results:

Demonstrated sequential release of prequantified lyophilized reagents from the multilayer tablet.
Successfully detected a nucleic acid target (tpp47 gene) and nitrite ions in aqueous samples autonomously.
Confirmed controlled dissolution rates of tablet layers for precise reagent delivery.

Conclusions:

The developed multilayer tablet system effectively automates sequential reagent introduction for multistep assays.
This technology significantly simplifies complex diagnostic procedures, enhancing usability for lay users.
The system holds potential for point-of-care diagnostics and reducing reliance on skilled personnel.