T. Photomicrograph gallery

Imaging Adipose Innervation

Fig. 1: Peripheral nerves lining a blood vessel residing in a mouse inguinal subcutaneous white adipose tissue (scWAT) depot. Stained with the pan-neuronal marker Beta III Tubulin. Visualized as a Z-maximum intensity projection that has been height-color-coded (purple close to surface, red at deeper level).

Micrograph captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 63X oil immersion objective. Post processing performed with LASX software and FIJI.

Photos on this page taken by Jake Willows, a Research Associate in the Townsend Lab at OSU (and previously a 4+1 Masters and Capstone student in our lab at UMaine). Jake has optimized techniques and made numerous novel discoveries when imaging the nerves in adipose tissue. (Learn more at Willows et al. 2021 on BioRxiv or at the lab’s protocols.io site)

Sympathetic Nerve Montage 10X

Fig. 4: Sympathetic Nerves in inguinal subcutaneous white adipose tissue are labeled with Tyrosine Hydroxylase (red). Visualized as a Z-maximum intensity projection. Micrograph captured on Nikon A1Rsi confocal microscope with a 10X dry objective. Post processing performed with NIS-Elements Advanced Research software and FIJI.

Sympathetic Nerve Montage 40x

Fig. 5: Small sympathetic neurites in subcutaneous white adipose depot visualized in 3 images using extended depth of field. Neurites labeled with tyrosine hydroxylase (red). Micrographs were capture on a Nikon E400 epifluorescence microscope with a 40X dry objective. Post processing was performed using NIS-Elements Basic Research software and FIJI.

Whole depot imaging of adipose nerves – Example 2

Fig. 7: Inguinal subcutaneous white adipose depots imaged as tiled z-stacks and max projected. Nerves were co-stained with 2 separate pan-neruonal markers Beta III Tubulin (green) and PGP9.5 (red). Nuclei labeled with DAPI (blue). Micrographs were captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 10X (top) and 5X (bottom) dry objectives. Post processing performed with LASX software and FIJI.

Digital Cross-Section of an Adipose Nerve Bundle

Fig. 2: Peripheral Nerve bundle in a mouse inguinal subcutaneous white adipose tissue (scWAT) depot. Stained with the pan-neuronal marker Beta III Tubulin (green). Left: nerve bundled visualized as a Z-maximum intensity projection. Right: the same nerve bundle captured in XZY to create a digital cross section.

Micrograph captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 63X oil immersion objective. Digital cross section has a 2.54X digital zoom applied.  Post processing performed with LASX software and FIJI.

Large nerve bundle in scWAT

Fig 3: Peripheral nerve bundle residing in an inguinal subcutaneous white adipose depot. Pan-neuronal marker Beta III Tubulin (green), pan-neuronal marker PGP9.5 (red), and nuclei marker DAPI (blue). Visualized as a Z-maximum intensity projection. Micrograph captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 63X oil immersion objective. Post processing performed with LASX software.

Whole depot imaging of adipose nerves – example 1

Fig. 6: Inguinal subcutaneous white adipose depots imaged as tiled z-stacks and max projected. Nerves marked with PGP9.5 (green) and blood vessels + sensory nerves marked with Isolectin-IB4 (red). Micrographs were captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 10X (top) and 5X (bottom) dry objectives. Post processing performed with LASX software and FIJI.

Whole Brain imaging – Example 1

Fig. 8: ~500um sagittal brain section from mouse optically cleared using iDISCO. Height color coded Z-maximum intensity projection emphasizing tissue structure through autofluorescence.

Micrograph captured on Leica TCS SP8 DLS point scanning confocal microscope using white light laser and 10X dry objective. Post processing performed with LASX software.