High-resolution TOF-SIMS imaging of eukaryotic cells preserved in a trehalose matrix.
A novel, trehalose-glycerol matrix was utilized to generate high-resolution, TOF-SIMS images of macrophages and glial cells. Viable cells incubated in 50 mM trehalose, then lyophilized in a 50 mM trehalose, 10-15% (w/w) glycerol rinse, are preserved and chemically profiled. These experiments demonstrate the utility of the disaccharide matrix as an efficient, cost-effective alternative to cryogenics for SIMS and other ultrahigh-vacuum (UHV) analyses of biological species. Cellular processes on oligodendrocytes and astrocytes, 1-3 mum in width, were well resolved for cells in the trehalose-glycerol matrix. The viscous cell matrixes were fractured and analyzed at room temperature and maintained their three-dimensional integrity under UHV. Images have been generated with a Au primary ion source near the static limit of 10(12) ions/cm2. Though these nucleated cells do not remain viable after desiccation, TOF-SIMS imaging and subsequent rehydration reveals structural and morphological preservation. Eliminating the inherent obstacles associated with cryogenic analysis opens the door to greater utility of SIMS as a bioanalytical tool, such as lipid mapping of single cells in the nervous system.
Parry
Shawn
S
Department of Chemistry, Pennsylvania State University, 209 Chemistry Building, University Park, Pennsylvania 16802, USA.
Winograd
Nicholas
N
D000818:Animals; D005057:Eukaryotic Cells / Q000648:ultrastructure*; D005612:Freeze Drying; D005990:Glycerol / Q000737:chemistry; D008264:Macrophages / Q000648:ultrastructure*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D009457:Neuroglia / Q000648:ultrastructure*; D051381:Rats; D014199:Trehalose / Q000737:chemistry*
D014199:Trehalose; D005990:Glycerol