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Related Experiment Video

Updated: May 23, 2026

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions

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Published on: January 30, 2018

Simultaneous quantitative analysis of three compounds using three-dimensional fluorescence spectra based on digital

Hong Lin Zhai¹, Zhi Jie Shan, Rui Na Li

¹State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou, 730000, People's Republic of China. zhaihl@163.com

Journal of Fluorescence

|April 6, 2012

Summary

A new quantitative approach using wavelet moment invariants for analyzing 3D fluorescence spectra accurately quantifies mixtures of Tryptophan, Tyrosine, and Phenylalanine, achieving high correlation coefficients.

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Area of Science:

Analytical Chemistry
Spectroscopy
Digital Image Processing

Background:

Digital image processing is widely used for qualitative analysis and classification.
Quantitative analysis of complex mixtures, especially with overlapping spectral peaks, remains challenging.

Purpose of the Study:

To propose a novel, simple, and quantitative approach for analyzing three-dimensional fluorescence spectra.
To accurately quantify amino acids (Tryptophan, Tyrosine, Phenylalanine) in mixture samples using digital image processing techniques.

Main Methods:

Calculation of wavelet moment invariants from digital grayscale images of 3D fluorescence spectra.
Development of linear models based on these invariants for quantitative analysis.
Application to mixtures of Tryptophan, Tyrosine, and Phenylalanine.

Main Results:

Achieved correlation coefficients (R²) greater than 0.99 for the linear models.
Demonstrated robustness against noise and background signals using statistical validation and cross-validation methods (leave-one-out, Jackknife).
Successfully quantified compounds even with overlapping spectral peaks.

Conclusions:

Wavelet moment invariants offer a robust and accurate method for the quantitative analysis of 3D fluorescence spectra.
The proposed approach is effective for analyzing complex mixtures, overcoming challenges posed by overlapping peaks.
This method provides a new pathway for spectral analysis in various scientific fields.