Mid-infrared metabolic imaging with vibrational probes.

Area of Science:

• Biophysics
• Chemical Biology
• Molecular Imaging

Background:

• Understanding cellular metabolism is crucial for deciphering physiological and pathological processes.
• Current in situ metabolic imaging tools lack the necessary throughput and specificity.
• There is a need for advanced techniques to visualize metabolic activities within biological systems.

Purpose of the Study:

• To develop a novel framework for in situ metabolic imaging using mid-infrared (MIR) microscopy.
• To introduce specifically designed IR-active vibrational probes for interrogating metabolic activities.
• To enable high-throughput metabolic profiling at single-cell and tissue levels.

Main Methods:

• Coupling high-information-throughput MIR microscopy with IR-active vibrational probes.
• Utilizing three categories of small vibrational tags: azide bonds, 13C-edited carbonyl bonds, and deuterium-labeled probes.
• Implementing broadband Fourier transform infrared microscopy and discrete frequency infrared microscopy with an extended spectral region (2,000–2,300 cm−1).

Main Results:

• Demonstrated a versatile framework for MIR metabolic imaging applicable to cells, small organisms, and mice.
• Successfully performed single-cell metabolic profiling, including heterogeneity characterization.
• Achieved large-area metabolic imaging at tissue and organ levels with rich spectral information.

Conclusions:

• The developed MIR metabolic imaging framework offers high information throughput for detailed metabolic analysis.
• This technique provides a powerful new tool for in situ metabolic research, advancing our understanding of biological processes.
• The study enables unprecedented insights into cellular and organismal metabolism through advanced vibrational probe and imaging strategies.