50 l nanosims  (CAMECA Inc)

Journal: Nature Communications

Article Title: Spatial patterns of hepatocyte glucose flux revealed by stable isotope tracing and multi-scale microscopy

doi: 10.1038/s41467-025-60994-w

Figure Lengend Snippet: A Illustration of the approach used to label freely moving and unanesthetized mice with [U- 13 C]-glucose for up to 4 hours using catheters followed by MIMS-EM and spatial analysis. Figure created using biorender.com. B Blood glucose levels measured from mice continuously infused with 15 or 40 mg/min/kg of total body mass for up to 4 hours. Data from n = 4-17 mice per group. C Expelled 13 CO 2 (in parts per million (ppm)) measured from the atmosphere of custom-made metabolic cages using gas mass spectrometers. 13 C glucose was infused for the first 240 minutes and replaced with 12 C glucose for an additional 120 minutes. AFE, atomic fractional enrichment. Data from n = 3 mice per group. D , E GC-MS analysis to determine the fractional 13 C enrichment of circulating glucose molecules and of secondary metabolites pyruvate, citrate, and alpha ketoglutarate (aKG) generated from glucose metabolism. Each dot represents an animal, n = 6 animals per time point. In B , data shown as ± standard deviation of the mean. In C , the shaded region indicates the data range of 13 CO 2 measurements. In D , E ), **** p < 0.001 and * p < 0.05 using One-way ANOVA with Kruskal-Wallis tests.

Article Snippet: Next, wafers containing the mapped samples were transferred to a MIMS microscope (50 L NanoSIMS, Cameca, France) for acquisition of multi-isotope maps ( 13 C, 12 C, 32 S, 14 N, and 31 P) as previously established , using the following MIMS image acquisition parameters: image size of 512×512 pixels, raster size of 30-to-40um 2 , at least three frames per raster with a 10 min acquisition time per frame using the beam adaptor D1-3 to yield a spatial resolution of ~80 nm in X-Y.

Techniques: Gas Chromatography-Mass Spectrometry, Generated, Standard Deviation

Journal: Nature Communications

Article Title: Spatial patterns of hepatocyte glucose flux revealed by stable isotope tracing and multi-scale microscopy

doi: 10.1038/s41467-025-60994-w

Figure Lengend Snippet: A , B Correlated 13 C-to- 12 C ( 13 C/ 12 C) ratiometric images acquired using MIMS and registered to scanning electron microscopy (SEM) of hepatocytes to create MIMS-EM maps. Data from mice continuously infused with [U- 13 C]-glucose at 40 mg/min/kg of [U- 13 C]-glucose for 1 or 4 hours. C Quantification of 13 C/ 12 C ratios in the biomass of mice shown in A , B . Data from n = 3 animals per time point. Each dot represents the mean 13 C enrichment in a ~40 um 2 liver section. D , E MIMS 13 C/ 12 C ratiometric images registered to hepatocyte SEM micrographs to create MIMS-EM maps. Data from mice continuously infused with15 or 40 mg/min/kg of total body mass for 2 hours. F Quantification of 13 C/ 12 C ratios in the biomass of mice from shown in D , E ). In A , B and D , E , magenta and orange colors represent lower and higher different levels of 13 C enrichment, respectively. Dotted magenta line marks the terrestrial background for 13 C. Data from n = 3 animals per time point. Each dot represents the mean 13 C enrichment in a ~40 um 2 liver section. G Cartoon illustration showing the single cell analysis of MIMS-EM data. Figure created using biorender.com. H Representative hepatocyte SEM, 13 C/ 12 C ratio, and overlay of several organelle segmentation masks, listed on the right. Data from a mouse continuously infused with 40 mg/min/kg of [U- 13 C]-glucose for 4 hours. I 13 C/ 12 C ratios of individual hepatocytes from mice continuously infused with 40 mg/min/kg of [U- 13 C]-glucose for 1 or 4 hours. Each color represents data from a different animal. J 13 C/ 12 C levels by type of organelle after 1 or 4 hours of 40 mg/min/kg [U- 13 C]-glucose infusion. K Relative fraction of hepatocyte cell area occupied by endoplasmic reticulum (ER), lipid droplets (LDs), glycogen, mitochondria, or nucleus. Data from n = 3-to-7 mice per group, from overnight fasted mice or from mice continuously infused with 40 mg/min/kg of [U- 13 C]-glucose for 1 or 4 hours. In I , J , data from n = 6-to-9 mice per group. In C and F ), a One-way ANOVA with a two-stage Benjamini, Krieger, and Yekuteli test was used, and p -values < 0.001 are shown. In J , K ), One-way ANOVA with Dunns test post-hoc where *** p < 0.001. In I – K , each dot represents a single cell. In (C and F-K), data shown as 95% of the confidence interval (C.I.). In (A), scale bar 2 microns; in D and H , 5 microns.

Article Snippet: Next, wafers containing the mapped samples were transferred to a MIMS microscope (50 L NanoSIMS, Cameca, France) for acquisition of multi-isotope maps ( 13 C, 12 C, 32 S, 14 N, and 31 P) as previously established , using the following MIMS image acquisition parameters: image size of 512×512 pixels, raster size of 30-to-40um 2 , at least three frames per raster with a 10 min acquisition time per frame using the beam adaptor D1-3 to yield a spatial resolution of ~80 nm in X-Y.

Techniques: Electron Microscopy, Single-cell Analysis