Lipid heterogeneity between astrocytes and neurons revealed with single cell MALDI MS supervised by immunocytochemical classification

Transcriptomics characterizes cells based on their potential molecular repertoire whereas direct mass spectrometry (MS) provides information on the actual compounds present within cells. Single cell matrix assisted laser desorption/ionization (MALDI) MS can measure the chemical contents of individual cells but spectra are difficult to correlate to conventional cell types, limiting the metabolic information obtained. We present a protocol that combines MALDI MS with immunocytochemistry to assay over a thousand individual rat brain cells. The approach entwines the wealth of knowledge obtained by immunocytochemical profiling with mass spectral information on the predominant lipids within each cell. While many lipid species showed a high degree of similarity between neurons and astrocytes, seventeen significantly differed between the two cell types, including four phosphatidylethanolamines elevated in astrocytes and nine phosphatidylcholines in neurons.

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