Dual-wavelength digital holographic phase and fluorescence microscopy combining with Raman spectroscopy for...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Spectroscopy

Background:

• Accurate detection and analysis of biomolecules are crucial for diagnostics and research.
• Existing immunoassay methods can suffer from crosstalk and limited encoding capacity.

Purpose of the Study:

• To develop a novel dual-channel encoding method for enhanced biomolecule detection and analysis.
• To combine dual-wavelength digital holographic phase microscopy (DW-DHPM) and Raman spectroscopy for a robust decoding platform.

Main Methods:

• Utilized Raman reporter molecules on micro-quartz pieces (MQPs) as microcarriers for suspension arrays (SA).
• Employed DW-DHPM and Raman spectroscopy as decoding platforms, with fluorescence microscopy for quantification.
• Implemented a dual-channel encoding strategy to avoid immunoassay crosstalk.

Main Results:

• Demonstrated accurate and stable decoding in multiplexed immunoassay experiments.
• Achieved high-sensitivity quantitative analysis of targets through concentration gradient experiments.
• Showcased expanded encoding capacity due to the combination of two encoding methods.

Conclusions:

• The developed DW-DHPFM technique offers a promising approach for sensitive and specific biomolecule detection.
• The dual-channel encoding method effectively minimizes crosstalk and enhances multiplexing capabilities.
• This integrated platform provides a powerful tool for advanced biochemical analysis.