Journal: The Journal of Neuroscience

Article Title: Actin-Cytoskeleton- and Rock-Mediated INM Are Required for Photoreceptor Regeneration in the Adult Zebrafish Retina

doi: 10.1523/JNEUROSCI.5005-14.2015

Figure Lengend Snippet: Live-cell imaging of INM in adult zebrafish retinal explants. A, Schematic of a retinal cross-section. The black arrows indicate the z-planes at the level of the ONL (Ba–Bk) and Müller glia soma (Ba′–Bk′). B, Time-lapse image series of retinal explant cultures from adult albino Tg[gfap:EGFP]nt11 zebrafish at 48 h of light treatment acquired by multiphoton microscopy showing that Müller glia divide in the ONL (arrows) and that their division plane is horizontal to the surface of the epithelium. Arrowheads indicate a Müller glia present in the ONL at the onset of image acquisition that displayed hallmarks of mitosis but did not divide during the 2 h recording. Scale bar, 20 μm. C–E, Histograms depicting the percentage of cells undergoing INM (C), the location of mitosis (D), and the division plane (E). F, Time-lapse image series of a 3D reconstruction of the z-series for the cell surrounded by a square in Bc. Arrows indicate the position of the cell soma as it migrates apically to the ONL and returns basally. Scale bar, 20 μm. G, H, Time-lapse image series of a 3D reconstruction of retinal cultures from adult Tg[gfap:nEGFP]mi2004 zebrafish giving an example of cells dividing horizontally (G) or vertically (H). Arrowheads point to the basal region of the nucleus or the soma after nuclear membrane breakdown. Stars indicate the time point of nuclear membrane breakdown. Scale bars in G, H, 10 μm. I, Graph depicting the distance the nucleus (F0) displayed in G and its daughter nuclei (D1, D2) traveled in relation to the basal INL position throughout the time lapse. Bars above the trace indicate the time period used to calculate the velocities of apical (va) and basal migration (vb). J, Histogram displaying the velocities of apical and basal nuclear migration. Images are representative from three independent retinal cultures. Data are shown as mean ± SE, nTg[gfap:EGFP] = 51 cells, nTg[gfap:nGFP] = 80 cells from three different retinal explants each. GCL, Ganglion cell layer.

Article Snippet: Live-cell imaging was performed using an inverted Nikon A1 Multiphoton microscope equipped with a 40× Apo long distance water-immersion objective (NA 1.15).

Techniques: Live Cell Imaging, Microscopy, Migration

Journal: Science advances

Article Title: Temporally resolved proteomics identifies nidogen-2 as a cotarget in pancreatic cancer that modulates fibrosis and therapy response.

doi: 10.1126/sciadv.adl1197

Figure Lengend Snippet: Fig. 5. NID2 reduction in CAFs leads to vascular changes in subcutaneous model under gemcitabine/Abraxane treatment shown via intravital imaging and quantum dots. (A) Subcutaneous coinjection intravital imaging experiment using GFP-1 KRAB or B500 NID2 KRAB CAFs (75%) with eGFP-tagged cancer cells (25%). Quantum dots in- jected via tail vein. Mice were treated twice weekly with gemcitabine/Abraxane, beginning day 11. n = 7 GFP-1 KRAB mice and n = 8 B500 NID2 KRAB mice. (B) Representative images for GFP-1 KRAB and B500 NID2 KRAB tumors imaged live via multiphoton intravital microscopy. SHG of fibrillar collagen (purple), eGFP cancer cells (green), vasculature (quantum dots; red), Imaris surface of blood vessels (red), and merged image. Scale bars, 100 μm. (C) Representative images of the vasculature of GFP-1 KRAB and B500 NID2 KRAB analyzed using VesselVio. Vessel skeletonization (black skeleton; top), skeletonization with 3D vessel rendering (black skeleton with red rendering; middle), and 3D vessel rendering alone (red rendering; bottom). Scale bars, 100 μm. (D) Quantification of total vessel volume (normalized to z depth of image in micrometers). Welch’s t test, *P < 0.05. (E) Quantification of total vessel surface area (normalized to z depth of image in micrometers). Welch’s t test, *P < 0.05. (F) Quantification of mean vessel radius (in micrometers) for GFP-1 KRAB (purple) and B500 NID2 KRAB (blue) live tumors. Welch’s t test, *P < 0.05. All data represented as means ± SEM. Schematics were created with Biorender.com.

Article Snippet: Images were acquired using a commercially available Leica STELLARIS 8 FALCON DIVE Multiphoton inverted microscope (Garvan ACRF INCITe Centre) with a Leica HC FLUOTAR L 25×/0.95 W VISIR powered by Spectra Physics Insight X3 single and MaiTai eHP Deep See and DMI8 inverted microscope’s external Leica 4Tune spectral Hybrid detectors (Leica Microsystems).

Techniques: Imaging, Intravital Microscopy