Journal: eLife

Article Title: A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids

doi: 10.7554/eLife.52904

Figure Lengend Snippet:

Article Snippet: Commercial assay or kit , Biomark 48.48integrated fluidic circuit Delta Gene assay , Fluidigm , Cat#:101–0348 , Complete bundle for 10 assays.

Techniques: Sequencing, Gene Assay, Viability Assay, Catecholamine ELISA, Software

Journal: Scientific Reports

Article Title: Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro

doi: 10.1038/s41598-018-24646-y

Figure Lengend Snippet: Schematic overview of the method described in this study. In ( A ) patterning of Dll4 by micro contact printing of the Dll4-fc immobilized to pink fluorescent beads in parallel lines with defined width and spacing. In ( B ) seeding of HUVECs through microfluidic channels perpendicular on the patterned Dll4. In ( C ) an overview of the HUVECs in (vessel mimic mimicking) monolayers perpendicular on top of the patterned Dll4 after removal of the microfluidic channels. ( D ) Hypothesized sprouting of the HUVECs after 24 h culture in Matrigel enriched media. Images made with tools from Servier medical art by Servier.

Article Snippet: 50 µl (10 µg/ml) of active human Dll4 protein fragment ab108557 (Abcam) was incubated with 100 µl (0.1% w/v) protein G fluorescent particles (Spherotech, purple, 0.4–0.6 µm) for 1 h at RT, resulting in a maximum of 0.5 µg of Dll4 ligand immobilized to the beads per sample.

Techniques:

Journal: Scientific Reports

Article Title: Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro

doi: 10.1038/s41598-018-24646-y

Figure Lengend Snippet: Fold change in gene expression of Ephrin B2 en Flt4 (VEGFR3). The experimental conditions are indicated below the graphs and in the legend adjacent. The condition of Beads with or without attached Dll4 underneath a gelatin coating is used here as well as the rest of this study. DMSO was used as a vehicle control for DAPT. Experiment in triple, qPCR was repeated 3 times. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: 50 µl (10 µg/ml) of active human Dll4 protein fragment ab108557 (Abcam) was incubated with 100 µl (0.1% w/v) protein G fluorescent particles (Spherotech, purple, 0.4–0.6 µm) for 1 h at RT, resulting in a maximum of 0.5 µg of Dll4 ligand immobilized to the beads per sample.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro

doi: 10.1038/s41598-018-24646-y

Figure Lengend Snippet: Microscopic immunofluorescent images of labelled sprouting HUVECs, in Matrigel enriched media, on top of gelatin coated glass (NoPrint, top row). Sprouting HUVECS in Matrigel enriched media on top of a line pattern of fluorescent IgG beads coated with gelatin (BeadOnly, middle row) or on a Dll4 ligand immobilized with fluorescent IgG beads line pattern coated with gelatin (BeadDll4, bottom row). First three columns contain intensity values separately. The last column contains the merged image with in green; actin stained with phalloidin, in pink; fluorescent IgG beads, and in blue; nuclei stained with DAPI. Scale bar represents 250 µm.

Article Snippet: 50 µl (10 µg/ml) of active human Dll4 protein fragment ab108557 (Abcam) was incubated with 100 µl (0.1% w/v) protein G fluorescent particles (Spherotech, purple, 0.4–0.6 µm) for 1 h at RT, resulting in a maximum of 0.5 µg of Dll4 ligand immobilized to the beads per sample.

Techniques: Staining

Journal: Scientific Reports

Article Title: Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro

doi: 10.1038/s41598-018-24646-y

Figure Lengend Snippet: Efficiency of controlled sprouting analysed with ( a ) 89 ROIs of bead-only condition and 98 ROIs of bead-Dll4 condition. ( b ) The reproducibility of the bead-Dll4 chips resulting from the Ecs calculated with 60 ROIs of Bead-Dll4 sample B and 71 ROIs of bead-Dll4 sample (c). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: 50 µl (10 µg/ml) of active human Dll4 protein fragment ab108557 (Abcam) was incubated with 100 µl (0.1% w/v) protein G fluorescent particles (Spherotech, purple, 0.4–0.6 µm) for 1 h at RT, resulting in a maximum of 0.5 µg of Dll4 ligand immobilized to the beads per sample.

Techniques:

Journal: Scientific Reports

Article Title: Spatial patterning of the Notch ligand Dll4 controls endothelial sprouting in vitro

doi: 10.1038/s41598-018-24646-y

Figure Lengend Snippet: Patterned Dll4 lines result in unidirectional endothelial sprouting. Direction of 5 × 5 pixel matrices was calculated for all ROI of either side of the vessel mimicking monolayer in one microscopy image, using the Image J plugin ‘Directionality’. ( a ) The dispersion of the fitted graph through the histograms in degrees. ( b ) The correctness of this fit through the direction histograms. *p < 0.05, **p < 0.01 ***p < 0.001, ****p < 0.0001. NP n = 15, BO n = 21, BD-A n = 23, BD-B n = 13, BD-C = 22. ( c ) Histogram of the directions found in the no print condition ( d ) for the bead-only control condition and ( e ) for the patterned Dll4 condition, with similar chips BeadDll4-B and C ( f and g ).

Article Snippet: 50 µl (10 µg/ml) of active human Dll4 protein fragment ab108557 (Abcam) was incubated with 100 µl (0.1% w/v) protein G fluorescent particles (Spherotech, purple, 0.4–0.6 µm) for 1 h at RT, resulting in a maximum of 0.5 µg of Dll4 ligand immobilized to the beads per sample.

Techniques: Microscopy