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Updated: May 9, 2025
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Autofluorescence Imaging to Evaluate Cellular Metabolism
Published on: November 15, 2021
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Exploring cellular metabolic kinetics through spectroscopic analysis of extracellular environments
Zohreh Mirveis1, Nitin Patil1, Hugh J Byrne2
1FOCAS Research Institute, TU Dublin, City Campus, Camden Row, Dublin 8, Ireland; School of Physics, Optometric and Clinical Sciences, TU Dublin, City Campus, Grangegorman, Dublin 7, Ireland.
Summary
Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy can analyze low metabolite concentrations. Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) effectively tracked glycolysis but struggled with complex mixtures containing glutamine.
Area of Science:
- Biochemistry and Spectroscopy
- Metabolic Pathway Analysis
- Cancer Metabolism Research
Background:
- Metabolic pathways like glycolysis and glutaminolysis are crucial in diseases such as cancer.
- Analyzing low concentrations of metabolites in extracellular media is essential for understanding metabolic shifts.
- Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy offers a potential method for such analyses.
Purpose of the Study:
- To explore Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy for quantifying low metabolite concentrations.
- To apply Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) for data mining metabolic pathway kinetics.
- To investigate the impact of glutamine on the analysis of glycolysis.
Main Methods:
- Utilizing ATR-FTIR spectroscopy to measure glucose, glutamine, and lactate concentrations in extracellular media.
- Employing sample extraction and drying on an ATR crystal for spectral analysis.
- Applying MCR-ALS to deconvolute time-series spectroscopic data of cellular metabolism.
Main Results:
- ATR-FTIR successfully measured low concentrations of glucose, glutamine, and lactate with strong absorbance-concentration correlations.
- MCR-ALS resolved glucose and lactate from glycolysis data, indicating a consumption rate of 0.0016 min⁻¹.
- MCR-ALS faced limitations in deconvoluting complex mixtures when glutamine overlapped with lactate signals.
Conclusions:
- ATR-FTIR spectroscopy is effective for analyzing low concentrations of key metabolites in metabolic studies.
- MCR-ALS can successfully model simple metabolic pathways like glycolysis but has constraints with complex mixtures.
- Further method development is needed to address spectral overlap challenges in multi-metabolite analyses.
