Spatial biology using single-cell mass spectrometry imaging and integrated microscopy.

Area of Science:

• Spatial biology
• Mass spectrometry imaging
• Cellular analysis

Background:

• Spatial biology analyzes cells in their microenvironment, including morphology and molecular expression.
• Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) provides spatial lipid and metabolic profiles.
• Integrating MALDI-MSI with other methods at the single-cell level is challenging due to co-registration and resolution issues.

Purpose of the Study:

• To develop a MALDI-MSI based method for integrated spatial analysis.
• To enable coupled (sub-)cellular investigation of the same sample using multiple modalities.
• To combine lipid profiling with morphological and protein expression data at the single-cell level.

Main Methods:

• Developed a MALDI-MSI method integrating in-source bright-field and fluorescence microscopy.
• Applied the method to cell culture and tissue samples.
• Enabled coupled analysis of lipid profiles and cellular features.

Main Results:

• Visualized intracellular lipid distributions in macrophages during phagocytosis.
• Revealed heterogeneity of lipid profiles in tumor-infiltrating neutrophils.
• Correlated lipid profiles with individual microenvironments.

Conclusions:

• The developed method allows for coupled (sub-)cellular investigation of lipid profiles and morphology.
• This approach provides a powerful tool for single-cell level analysis in cell biology.
• Enables deeper understanding of cellular functions within their native microenvironments.