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A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting

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Electrochemiluminescence Amplification in Bead-Based Assays Induced by a Freely Diffusing Iridium(III) Complex.

Emily Kerr¹, Sara Knezevic², Paul S Francis³

¹Institute for Frontier Materials, Deakin University, Geelong, Victoria 3220, Australia.

|January 26, 2023

Summary

This study introduces a new method to boost electrochemiluminescence (ECL) signals in assays by adding an iridium complex. This enhancement improves sensitivity and opens new possibilities for bioanalytical detection and microscopy.

Keywords:

bead-based assays electrochemiluminescence electrochemistry heterogeneous assays imaging

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Area of Science:

Electrochemistry
Analytical Chemistry
Materials Science

Background:

Heterogeneous electrochemiluminescence (ECL) assays commonly use tris(2,2'-bipyridine)ruthenium(II) ([Ru(bpy)₃]²⁺) and tri-n-propylamine.
This system is crucial for many ECL sensing applications and bead-based immunoassays.
Enhancing the ECL signal is vital for improving assay sensitivity and performance.

Purpose of the Study:

To develop a novel method for enhancing conventional heterogeneous ECL assays.
To investigate the effect of incorporating a second metal coordination complex, [Ir(sppy)₃]^3-, into the ECL system.
To analyze the spatial distribution of ECL emission using ECL microscopy.

Main Methods:

Utilized ECL microscopy to map emission from [Ru(bpy)₃]²⁺ labels and [Ir(sppy)₃]^3-.
Incorporated the iridium complex [Ir(sppy)₃]^3- into a standard ECL assay system.
Studied the enhancement of ECL signals from immobilized [Ru(bpy)₃]²⁺ labels on polystyrene beads.

Main Results:

The addition of [Ir(sppy)₃]^3- significantly enhanced the ECL signal from [Ru(bpy)₃]²⁺ labels (70.9-fold at 0.9 V, 2.9-fold at 1.2 V vs Ag/AgCl).
ECL microscopy visualized distinct spatial distributions of emission from both the ruthenium label and the iridium complex.
The enhancement was particularly pronounced at low oxidation potentials.

Conclusions:

The [Ir(sppy)₃]^3--mediated approach offers a substantial improvement for heterogeneous ECL assays.
This method can amplify ECL signals, enabling new bioanalytical detection strategies.
The enhancement may reduce electrode passivation and side reactions, improving assay reliability.