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Fluorescence detection methods for microfluidic droplet platforms

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Ultrasensitive Qbeta phage analysis using fluorescence correlation spectroscopy on an optofluidic chip.

M I Rudenko¹, S Kühn, E J Lunt

¹School of Engineering, University of California, Santa Cruz, Santa Cruz, CA 95064, USA. mrudenko@soe.ucsc.edu

Biosensors & Bioelectronics

|May 16, 2009

Summary

We developed an optofluidic biosensor for single Qbeta bacteriophage detection and analysis in their natural liquid state. This technology allows for virus characterization without immobilization, showing potential for portable virus discrimination.

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Multicolor Fluorescence Detection for Droplet Microfluidics Using Optical Fibers

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Area of Science:

Optofluidics
Nanobiotechnology
Virology

Background:

Accurate detection and analysis of individual viruses are crucial for understanding viral behavior and developing diagnostics.
Existing methods often require virus immobilization, which can alter their natural state and introduce artifacts.

Purpose of the Study:

To demonstrate a novel optofluidic biosensor for label-free, single-virus level detection and analysis of Qbeta bacteriophage.
To analyze virus behavior, including diffusion, aggregation, and flow, in a native liquid environment.

Main Methods:

Utilized an integrated optofluidic biosensor on a silicon chip.
Employed fluorescence correlation spectroscopy (FCS) to analyze fluorescence signals from individual viruses.
Sensed viruses in their natural liquid environment without immobilization.

Main Results:

Successfully detected and analyzed individual Qbeta bacteriophages (attogram mass range) in real-time.
Determined the diffusion coefficient to be 15.90±1.50 µm²/s, consistent with literature values.
Observed virus aggregation and disintegration, and measured flow velocities between 60-300 µm/s.

Conclusions:

The developed optofluidic biosensor enables sensitive, label-free detection and analysis of viruses at the single-particle level.
The system operates without virus immobilization, preserving their native state.
This technology holds significant potential for developing inexpensive, portable sensors for discriminating between viruses of different sizes.