Description

Topographical regulation of keratinocyte migration. 2D, 3D, and 1D fibrillar migration of human epidermal keratinocytes. 2D matrices were constructed by uniform coating with extracellular matrix (ECM). 3D surfaces were constructed according to Science. 294: 1708–1712 (2001). 1D surfaces were made by using a 2 photon confocal microscope to pattern a polyvinyl alchohol (PVA) coated coverslip. Movie is video 4 from JCB 2009, 184:481-490

Technical Details

Following gluteraldehyde quenching, ECM proteins were absorbed onto the PVA surface. Videos were recorded on a Axiovert 135TV (Carl Zeiss, Inc.) with a charge-coupled device camera (ORCA II ER; Hamamatsu Photonics). MetaMorph imaging software was used to acquire images and control all hardware. A custom environmental chamber (Lucite) enclosed both time-lapse and TIRF microscopes and maintained cells at 37°C with 10% CO2. Images were collected every 4 min and shown at 15 frames per second.