Principle of the cell spot microarray (CSMA) method. (A) CSMA work flow: siRNA samples in a printing solution containing transfection lipid and extra-cellular matrix proteins are robotically printed on a hydrophobic polystyrene surface. A suspension of adherent cells is allowed to adhere onto the array spots, followed by washing un-adhered cells off, leaving adhered cells only to printed array positions. After reverse transfection for a selected time, the arrays are stained using e.g. traditional multi-label immunostaining protocols for high content image analysis of multiple parameters. (B) CSMA method allows production of high density patterned cell arrays. Left panel displays a microarray scanned view of 200 μm cell spots with 500 μm spot spacing. Scale bar 0,5 mm. Right panel: Due to the spatially confined layout of the spots, automated imaging and segmentation of cells on CSMA spots can be performed using automated image analysis software. Scale bar 200 μm. (C) Microscopic image and microarray scanned view of 200 μm CSMA spots (Top left panel;i) and 400 μm spots (Top right;ii) of PC-3 cells stained for DNA (blue) and F-Actin (green) after 48 h culture. Microscopic image and microarray scanned view of 200 μm CSMA spots (Bottom;iii) of primary prostate stromal cells cultured for 48 h and stained for DNA (blue) and F-Actin (green). Scale bar 900 μm. (D) Phase contrast microscopic images from a timelapse series of PC-3 cells cultured on 200 μm array spots for 72 h.
Rantala et al. BMC Genomics 2011 12:162 doi:10.1186/1471-2164-12-162