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Related Experiment Video

Updated: May 28, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

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Published on: December 9, 2013

Parallel-scan based microarray imager capable of simultaneous surface plasmon resonance and hyperspectral

Zhiyi Liu¹, Lei Yang, Le Liu

¹Laboratory of Optical Imaging and Sensing, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, PR China.

Biosensors & Bioelectronics

|October 15, 2011

Summary

This study introduces a novel combined technique for simultaneous surface plasmon resonance (SPR) sensing and hyperspectral fluorescence imaging. This method enhances microarray analysis by providing comprehensive, dual-validation data for biological events.

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Area of Science:

Biotechnology
Analytical Chemistry
Biosensing

Background:

Microarray technology necessitates advanced detection methods for complex biomolecular interactions.
Simultaneous analysis of multiple factors influencing these interactions is challenging with single techniques.
Understanding the interplay of factors is crucial for accurate biological event interpretation.

Purpose of the Study:

To develop a combined technique for simultaneous angle-interrogation surface plasmon resonance (SPR) sensing and hyperspectral fluorescence imaging.
To enable comprehensive, two-dimensional imaging and analysis of biological events on microarray chips.
To validate experimental results and exclude false positives/negatives through dual-detection confirmation.

Main Methods:

A tandem system integrating angle-interrogation SPR sensing with hyperspectral fluorescence imaging was developed.
The system provides simultaneous refractive index information (SPR) and physicochemical properties (fluorescence).
Two model experiments involving DNA hybridization and biotin-avidin interactions were conducted.

Main Results:

The SPR sensing achieved a refractive index resolution of 3.86×10⁻⁶ RIU and a detection limit of 10⁴ cfu/ml for E. coli.
Hyperspectral fluorescence imaging demonstrated a resolving power of approximately 20 μm and sensitivity of 0.61 fluors/μm².
The combined technique successfully detected DNA hybridization and biotin-avidin interactions, validating its biomedical applicability.

Conclusions:

The proposed tandem SPR and fluorescence imaging technique offers a powerful tool for comprehensive microarray analysis.
Simultaneous dual detection enhances accuracy and reliability by confirming experimental findings.
This integrated approach significantly advances the capabilities for studying complex biological events in biomedical applications.