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Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

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Fluorescent Lateral Flow Immunoassay Based on Quantum Dots Nanobeads.

Lingzhi Fan¹, Yue Luo², Wannian Yan¹

¹Department of Central Laboratory, Shanghai Skin Disease Hospital, School of Medicine, Tongji University.

Journal of Visualized Experiments : Jove

|July 15, 2024

Summary

Researchers developed quantum dot nanobeads (QDNB) for enhanced sensitivity in disease biomarker detection. This novel approach improves lateral flow immunoassay performance for faster and more accurate diagnostics.

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Author Spotlight: High-Quality Quantum Dot Nanobeads for Sensitive Fluorescent Lateral Flow Immunoassays

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Published on: June 28, 2024

Area of Science:

Biotechnology
Nanotechnology
Analytical Chemistry

Background:

Quantum dots (semiconductor nanocrystals) are fluorescent labels used in bioimaging and sensing.
Conventional quantum dot-antibody conjugates (~10 nm) show limited sensitivity in trace disease marker detection via lateral flow immunoassays (LFIA).
Laborious purification methods hinder the development of sensitive quantum dot-based diagnostic tools.

Purpose of the Study:

To develop a novel method for preparing quantum dot nanobeads (QDNB) for improved LFIA sensitivity.
To fabricate a fluorescent LFIA using QDNB for rapid disease biomarker detection.
To demonstrate the enhanced sensitivity and efficiency of QDNB-based immunoassays.

Main Methods:

A one-step emulsion evaporation method was employed to synthesize QDNB.
QDNB were conjugated with antibodies to create QDNB-antibody conjugates.
A fluorescent lateral flow immunoassay was developed using QDNB-antibody conjugates for C-reactive protein (CRP) detection.

Main Results:

The developed QDNB-based LFIA demonstrated significantly higher sensitivity compared to single quantum dot nanoparticles.
Signal amplification was achieved by encapsulating hundreds of quantum dots within each nanobead.
The larger size of QDNBs simplified antibody conjugation through easier centrifugation.
The assay successfully detected CRP concentration within 15 minutes, with results assessable qualitatively (UV light) and quantitatively (fluorescent reader).

Conclusions:

The one-step synthesis of QDNB offers an efficient alternative for preparing sensitive immunoassay labels.
QDNB-based fluorescent LFIA provides a rapid, sensitive, and quantitative method for disease biomarker detection.
This technology holds promise for improving point-of-care diagnostics and early disease screening.