HUVECs were seeded onto 50 mm imaging dishes (Ibidi, Martinsried, Germany). When the cells achieved 100% confluence, a straight lesion was made in the center of the monolayer using a sterile 200-μL pipette tip. The wells were subsequently washed twice with PBS to remove the dead cells and incubated with EBM, containing 1 mM melatonin (final concentration of ≤0.1% DMSO), 1 mM 3-indolacetic acid (final concentration of ≤0.1% DMSO), or vehicle controls (≤0.1% DMSO) for 4 h before being treated with VEGF (25 ng/mL) for 24 h. The wounds were photographed using phase contrast microscopy on an inverted microscope (Nikon, Tokyo, Japan). Initial and final wound sizes were determined using the Nis-Elements BR v.4.30.02 software (Nikon, Tokyo, Japan), and the difference was used to determine the migration distance using the following formula; initial wound size minus final wound size divided by two. Three independent experiments were carried out.
Nis elements br v 4
Manufactured by Nikon
Sourced in Japan
Nis-Elements BR v.4.30.02 is a software application for imaging and analysis. It provides tools for image acquisition, processing, and data management. The software supports a range of microscopy techniques and imaging hardware.
Automatically generated - may contain errors
4 protocols using nis elements br v 4
1
Melatonin and 3-indolacetic Acid Modulate HUVEC Migration
2
Serotonin and 5-Hydroxytryptophol Regulate HUVEC Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell migration assay was performed following the protocol previously described in Cerezo et al. [19 (link)]. The HUVECs were placed onto 50 mm imaging dishes (Ibidi, Martinsried, Germany). Once cells reached full confluence, the monolayer was scratched across the centre using a sterile 200 μL pipette tip. Subsequently, the cells were then rinsed twice with PBS to eliminate any dead cells. Serum-free medium containing 1 mM serotonin, 1 mM 5-hydroxytryptophol or vehicle controls were added (final concentration ≤ 0.1% DMSO) for 4 h prior to VEGF stimulation (25 ng/mL) over 24 h. A phase contrast microscope (Nikon, Tokyo, Japan) was used to photograph the wound-gaps. Nis-Elements BR v.4.30.02 software (Nikon, Tokyo, Japan) was utilised to measure the initial and final sizes of each wound. The following formula was used to quantify the migration distance: initial wound size minus final wound size divided by two. Two independent experiments were performed.
3
Visualizing Vascular Structure in Mouse Somatosensory Cortex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surgical procedures were performed as described above. Ringers’ solution containing fluorescein isothiocyanate-labeled serum albumin (FITC-BSA) was topically superfused over the somatosensory cortex for 30 min. The mouse was then transcardially perfused with 20 ml of phosphate-buffered saline (PBS) and 30 ml of 4% paraformaldehyde, and, after sacrificing the animal, the brain was removed and post-fixed in 4% paraformaldehyde overnight. The brain was sectioned in 50-µm-thick coronal slices through the perfusion site using a vibratome, washed in PBS, immunostained with Alexa 594-conjugated anti-smooth muscle α-actin (1:500, clone 1A4; Abcam, Paris, France) and mounted on a glass slide in a drop of Dako fluorescence mounting medium (Dako; Les Ulis, France). Stained sections were imaged with a Nikon Eclipse 80i microscope (Nikon; Champigny sur Marne, France); images were captured with an Andor Neo sCMOS camera and NIS Elements BR v 4.0 software (Nikon) using identical settings across compared groups.
4
Quantifying Cellular and Vascular Pathology
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!