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Fluorescence detection in capillary arrays based on galvanometer step scanning.

G Xue¹, E S Yeung

¹Ames Laboratory-USDOE and Department of Chemistry Iowa State University, USA.

Electrophoresis

|October 24, 2001

Summary

A novel galvanometer scanner enables efficient laser-induced fluorescence detection in 96-capillary arrays. This system achieves a low limit of detection for fluoresceins with minimal cross-talk, enhancing analytical capabilities.

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Area of Science:

Analytical Chemistry
Biophysical Chemistry
Instrumentation

Background:

Capillary array electrophoresis (CAE) is a powerful separation technique.
Laser-induced fluorescence (LIF) is a sensitive detection method for CAE.
Efficient scanning of multiple capillaries is crucial for high-throughput analysis.

Purpose of the Study:

To adapt a computer-controlled galvanometer scanner for scanning a focused laser beam across a 96-capillary array.
To enable laser-induced fluorescence detection with temporal sorting of signals.
To evaluate the detection limits and cross-talk of the developed system.

Main Methods:

Utilized a computer-controlled galvanometer scanner to direct a focused laser beam.
Employed a 96-capillary array for sample separation.

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Implemented a single photomultiplier tube with temporal signal sorting for capillary discrimination.

Measured the limit of detection for fluoresceins at a laser power of 5 mW and scan frequency of 4 Hz.

Main Results:

Achieved a limit of detection of 3 x 10(-11) M for fluoresceins (Signal-to-Noise ratio = 3).
Observed minimal cross-talk (0.2%) among capillaries.
Demonstrated efficient light utilization due to a high duty-cycle step scan.
Reduced stray light, leading to improved detection performance.

Conclusions:

The adapted galvanometer scanner provides an efficient and cost-effective solution for LIF detection in 96-capillary arrays.
The system offers high sensitivity, low cross-talk, and flexibility for various analytical applications.
Advantages include efficient light use, reduced stray light, system ruggedness, low cost, and independent excitation/emission paths.