Skip to main content
. 2021 Sep 11;24(10):103127. doi: 10.1016/j.isci.2021.103127

Figure 7.

Figure 7

Whole depot neurite density quantification and heatmap generation

(A) Female C57BL/6J mice, aged 14–16 weeks, were either cold exposed (5°C) (n = 6) or maintained at room temperature (n = 4) for 7 days. Axi-scWAT depots were whole mount processed and stained with PGP9.5 and imaged with 10X objective. To demonstrate the difference between true signal and tissue autofluorescence pixel intensity less than 55 was considered to be autofluorescence (gray) and pixel intensity 55 or greater was considered true staining (green).

(B) Images were Zmax projected, tiled, and thresholded and neurite density quantification was performed, compared across the cohort, and analyzed using a two-tailed Student’s ttest. Alpha level 0.05, error bars are SEMs.

(C) Neurite density per Zmax tile was calculated and used to generate a heatmap.

(D) Ing-scWAT depot was stained with IB4 to mark vasculature, imaged at 10X, and vascular density per Zmax tile was calculated and used to generate heatmap. Each tile is comprised of a 0.12 cm × 0.12 cm area (C and D). All imaging was performed with a Leica TCS SP8 DLS laser scanning confocal microscope. Scale bars are 2 mm (A).