Submicron 3-D mass spectrometry imaging reveals an asymmetric molecular distribution on chemotaxing cells [version 1; peer review: 2 approved]

Abstract

Background: Dictyostelium discoideum is a ~10 µm diameter unicellular eukaryote that lives on soil surfaces. When starved, D. discoideum cells aggregate into streams of cells in a process called chemotaxis. In this report, we studied D. discoideum cells during chemotaxis using 3D - mass spectrometry imaging (3D-MSI). Methods: The 3D-MSI consisted of the sequential generation of 2D molecular maps using burst alignment coupled to delayed extraction time-of flight secondary ion mass spectrometry (TOF-SIMS) combined with a soft sputtering beam to access the different layers. Results: Molecular maps with sub-cellular high spatial resolution (~300 nm) indicated the presence of ions at m/z = 221 and 236 at the front and sides, but reduced levels at the back, of cells moving toward of aggregation streams. The 3D-MSI also detected an ion at m/z = 240 at the edges and back, but reduced levels at the front, of aggregating cells. Other ions showed an even distribution across the cells. Conclusions: Together, these results demonstrate the utility of sub-micron MSI to study eukaryotic chemotaxis.

Keywords

• Dictyostelium
• chemoattraction
• mass spectrometry imaging
• aggregation
• molecular imaging
• eng