Elucidating time-resolved intracellular metabolic dynamics via label-free Raman microspectroscopy and 2D correlation...

Area of Science:

• Cellular metabolism
• Biophysics
• Spectroscopy

Background:

• Metabolic processes like glycolysis and glutaminolysis are crucial for cellular function and disease.
• Dysregulation of these pathways is linked to various disorders.
• Raman spectroscopy offers a non-invasive method to study cellular dynamics.

Purpose of the Study:

• To investigate cellular metabolic responses to glucose and glucose-glutamine using Raman spectroscopy and 2D-COS.
• To understand the specific role of glutamine in modulating metabolic dynamics.
• To validate the use of 2D-COS for analyzing complex metabolic data.

Main Methods:

• Cells were starved and then exposed to glucose or glucose-glutamine.
• Spectroscopic monitoring was performed at 15-minute intervals for up to 2 hours.
• Two-dimensional correlation spectroscopy (2D-COS) was applied to analyze Raman spectra and kinetic data.

Main Results:

• Simulated data confirmed 2D-COS reliability in tracking dynamic variables.
• Cells under glucose-only conditions showed synchronized metabolic responses with oscillations.
• Glucose-glutamine conditions exhibited accelerated and amplified metabolic variability, with increased correlation and nucleic acid band activity.

Conclusions:

• Raman 2D-COS effectively resolves intracellular metabolic dynamics from complex datasets.
• Glutamine significantly influences metabolic variability and synchronization.
• This technique holds promise for advancing diagnostics and therapeutic interventions in metabolic disorders.