Optimized, automated assays for HCS

This reliable, automated assay identifies angiogenic tubes (micro-capillaries) formed by endothelial cells and provides quantitative measurements related to angiogenic tube formation such as the number of tubes, their morphology and branching, and an angiogenic index (% of image area connected by tubes x10). This assay can be run with a single fluorescent label (e.g., phalloidin conjugate or Invitrogen CellMask stains for fixed cells) to identify tubes, or additional channels can be used to report cell nuclei or target-specific proteins.

Typical angiogenesis assay setup

Can be assayed in a fixed-cell or live-cell format with appropriate labels.

Imaging mode

  • Widefield or confocal
  • 5x magnification

Automatically measured properties

  • % connected tubes
  • Mean connected tube area
  • Mean connected width
  • Mean connected tube node spacing
  • Angiogenic index

Angiogenesis assay images

HUVEC cells, grown in Matrigel support and treated with angiogenic compounds and imaged at 5x magnification. (Left) Cell nuclei are stained with Invitrogen Molecular Probes Hoechst 33342 (shown in red) and the cytoskeleton (actin fibers) is stained using a phalloidin conjugate (shown in green/yellow; wide choice of labels). (Right) Angiogenesis features identified using Thermo Scientific HCS Studio Software. In a two-channel assay, connected tubes are automatically identified using a blue overlay, and branching nodes are displayed as pink spots. Multiple parameters can be automatically measured in additional detection channels and correlated with the identified tube structures.

Sample experimental data

Suramin serves as a potent inhibitor of angiogenesis. A dose-response for suramin concentration can be plotted against a variety of properties measured automatically using Thermo Scientific ArrayScan VTI with Thermo Scientific HCS Studio software and plotted using GraphPad Prism software.