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Immunofluorescence Microscopy

Immunofluorescence Microscopy

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.
The...

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Updated: Jun 3, 2026

Multiplexed Fluorescent Microarray for Human Salivary Protein Analysis Using Polymer Microspheres and Fiber-optic Bundles

Multiplexed Fluorescent Microarray for Human Salivary Protein Analysis Using Polymer Microspheres and Fiber-optic Bundles

Published on: October 10, 2013

Microfluidic pillar array sandwich immunofluorescence assay for ocular diagnostics.

James V Green¹, Dawei Sun, Ali Hafezi-Moghadam

¹Department of Chemical Engineering, Northeastern University, Boston, MA, 02115, USA.

Biomedical Microdevices

|April 5, 2011

Summary

This summary is machine-generated.

A novel microfluidic device improves diagnosis of eye diseases like uveitis and primary intraocular lymphoma (PIOL). This sensitive assay detects critical biomarkers in ocular fluids, surpassing current limitations for accurate patient stratification.

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Published on: November 13, 2017

Area of Science:

Ophthalmology
Biomedical Engineering
Analytical Chemistry

Background:

Uveitis and primary intraocular lymphoma (PIOL) present diagnostic challenges due to similar symptoms and low sensitivity of current diagnostic methods.
Existing techniques for analyzing ocular fluid biomarkers have a diagnostic yield below 20% due to inadequate sensitivity.
Accurate diagnosis is crucial for effective treatment and improved patient outcomes in these distinct ocular conditions.

Purpose of the Study:

To develop and validate a novel microfluidic device for sensitive detection of biomarkers in ocular samples.
To overcome the limitations of current diagnostic techniques for uveitis and PIOL.
To enable accurate quantification of cytokine concentrations in ocular fluids for improved disease diagnosis and management.

Main Methods:

A simple sandwich immunofluorescence assay (sIFA) microfluidic device was designed and fabricated.
The device utilizes antibody-coated pillars for capture of target molecules, followed by detection with biotinylated antibodies and fluorescent avidin.
Cytokine concentrations were measured in aqueous humor samples from rats with induced uveitis and human cataract patients.

Main Results:

The sIFA device demonstrated high sensitivity, achieving single-digit femtomolar detection limits for cytokines.
Results from ocular fluid samples correlated well with conventional protein quantification techniques.
The assay surpassed the detection limits of current state-of-the-art immunoassay techniques for human samples.

Conclusions:

The developed sIFA microfluidic device offers a sensitive and cost-effective alternative for diagnosing ocular diseases.
Its high sensitivity and potential for automation facilitate translation to clinical settings for improved patient diagnosis.
This technology enables precise mapping of cytokine profiles in vitreous biopsies, advancing diagnostic capabilities in ophthalmology.