Widefield surface plasmon resonance microscope: A novel biosensor study of cell attachment to micropatterned substrates

In this study, Human adult, low Ca2+, high Temperature (HaCaTs) Keratinocytes were cultured on microcontact printed fibronectin repeat gratings of 1.8, 3.8, 5, 12.5 and 25μm for 24 hours. The cells were then fixed with 0.1% Glutaraldehyde and dehydrated in serial alcohol [1]. The alignment of the cells were then measured, where 0º represents 100% alignment to the pattern, in order to identify those features that promoted the highest degree of cell alignment. From the quantitative analysis it became clear that HaCaTs cells align most readily to the 12.5μm pattern. A 12.5μm stamp was therefore used to stamp pattern fibronectin on to prefabricated Au/Cr/glass surface plasmon substrates. HaCaTs cells were cultured on the substrates for 24 hours and imaged with the “Widefield Surface Plasmon Resonance” (WSPR) microscope [1]. The WSPR system enables the examination of nanometric interfacial interactions of HaCaTs on patterned and un-patterned surface at lateral resolution down to 500nm. Our preliminary results demonstrate that the WSPR microscope is capable of imaging the cell surface interface and as such will prove to be a very useful tool in understanding the processes involved in cell guidance and wound repair.