Probing the Subcellular Localization of Hopanoid Lipids in Bacteria Using NanoSIMS
Figure 5
Analysis of BHT localization in N. punctiforme cells.
NanoSIMS images of 1H− (A) and 2H− (B) ion count rate and a non-quantitative overlay (C) of the raster images depicting their relative localizations with 1H− ions shown in green and 2H− ions shown in red. The thin white circles in C denote the regions of interest (numbered 1–8) selected for the pooled analysis of data in panel D. The white arrow indicates that region of interest 4 is within region of interest 2. (D) Plot depicting the δ13C and δ2H values for the regions of interest (numbered blue circles) and all image pixels containing greater than 100 cts s−1 1H− ions, which encompasses all biomass in the image (blue square). Additional regions of interest points shown as are data taken from other ion images (unnumbered blue circles). The line is an ordinary least squares regression of interest shown in C. (E) A white dashed line indicates the region enlarged in panel F. (F) The white dashed line indicates a region of interest along a filament for the isotopic profile shown in G. (G) δ2H values along the filament highlight a periodicity with labeled BHT concentrated at the junctions between cells. All scale bars are 5 µm. Error bars in (G) reflect the standard error of the total number of ions. Images shown are representative of 5 fields of view and 3 independent biological experiments.