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The major problem of Digital Holographic Microscopy (DHM) long term live cell imaging is that over time most of the tracked cells move out of the image area and other ones move in. Therefore, most of the cells are lost for the evaluation of individual cellular processes. Here, we present an effective solution for this crucial problem of long-term microscopic live cell analysis. We have generated functionalized slides containing areas of 250 μm per 200 μm. These micropatterned biointerfaces consist of passivating polyaclrylamide brushes (PAAm). Inner areas are backfilled with octadecanthiol (ODT), which allows cell attachment. The fouling properties of these surfaces are highly controllable and therefore the defined areas designed for the size our microscopic image areas were effective in keeping all cells inside the rectangles over the selected imaging period.
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Sarah Mues, Inga Lilge, Holger Schönherr, Björn Kemper, Jürgen Schnekenburger, "Micro patterned surfaces: an effective tool for long term digital holographic microscopy cell imaging," Proc. SPIE 10074, Quantitative Phase Imaging III, 1007410 (21 February 2017); https://doi.org/10.1117/12.2253280