Autofluorescence profiles define depot-specific adipocyte phenotypes in perivascular and non-perivascular adipose tissue

Figures

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  Figure 1.. PVAT and non-PVAT adipocytes’ viability is maintained after enzymatic digestion. A) Quantitative assessment of adipocyte viability following enzymatic digestion. Viability is expressed as a percentage of live cells (acridine orange–AO+)/dead cells (propidium iodide–PI+) x 100 in different adipose depots: thoracic aortic PVAT (atPVAT), interscapular brown adipose tissue (BAT), mesenteric PVAT (mesPVAT), retroperitoneal adipose tissue (rpAT), abdominal PVAT (abPVAT), and subcutaneous adipose tissue (SCAT). Bars represent mean ± SEM. B) Representative CellometerTM Spectrum images showing live (AO+, green fluorescence) and dead (PI+, red fluorescence) cells from each adipose depot.

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  Figure 2.. PVAT and non-PVAT depots differ in adipocyte size and SSC profiles. A) Representative scatter plots showing the distribution of forward scatter (FSC; x-axis) and side scatter (SSC; y-axis) for all events and gating for adipocytes of combined data from atPVAT (orange) and BAT (teal), D) combined data from rpAT (yellow), mesPVAT (lime), and G) combined data from SCAT (red) and abPVAT (green). B, E, H) Histogram of events from adipo gate showing distribution of counts across FSC signature between PVAT and non-PVAT depots (left) and bar graph summarizing cell area in µm2 in mean ±SEM (right). C, F, I) Representative images obtained via the Attune CytPix demonstrating the characteristic multilocular phenotype in PVAT depots and their counterparts, varying from unilocular to multilocular adipocytes. White adipocytes present nuclei in the periphery of the cell. Scale bar: 10 µm.

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  Figure 3.. Autofluorescence profiles of PVAT depots and their non-PVAT counterparts. A) Quantitative comparison of median fluorescence intensity (MFI; y-axis) between PVAT and non-PVAT depots across blue (BL)-1, yellow (YL)-2, red (RL)-3, and violet (VL)-3 channels. Data are presented as median ± SE, different letters: p < 0.05. B) Scatter plots illustrating autofluorescence intensity in diverse filter channels across multiple blue (BL), yellow (YL), red (RL), violet (VL), and violet (VL) channels.