Fig. 1: Engineering ECM protein patterns with defined sizes by microcontact printing (µCP). | Nature Communications

Fig. 1: Engineering ECM protein patterns with defined sizes by microcontact printing (µCP).

From: Mammalian cells measure the extracellular matrix area and respond through switching the adhesion state

Fig. 1: Engineering ECM protein patterns with defined sizes by microcontact printing (µCP).

A Illustration of the production of polydimethylsiloxan (PDMS) stamps having micropillars of different diameters. First, a silicon wafer with arrays of circular holes with 10 μm, 8 μm, 5 μm, 3 μm, or 2 μm diameter is fabricated by photolithography. Then, PDMS is poured onto the wafer, cured, and subsequently the PDMS stamp (master) with micropillars is peeled off from the wafer. B Differential interference contrast (DIC) microscopy of the top layer of PDMS stamps with arrays of micropillars of different diameters. Scale bar, 20 μm. C µCP-based patterning of ECM proteins on glass surfaces. First, the PDMS stamp is coated (inked) with fluorescently labeled (Alexa fluor 555) ECM proteins (left), afterwards washed with PBS and deionized water to remove residues, then blow-dried by air (middle), and finally ECM proteins are printed on glass through physical contact (right). D Confocal microscopy images of circular collagen I (top) and fibronectin (bottom) patterns printed with PDMS stamps having arrays of micropillars of different diameters. Scale bars, 20 μm. E Fluorescence images of (I) Alexa fluor 555-labeled collagen I (top) or fibronectin (bottom) patterns printed on glass using PDMS micropillars having 10 µm diameter. (II) After printing of ECM proteins, the uncoated glass surfaces are passivated with FITC-labeled BSA (top) or Alexa fluor 488-labeled FNIII7-10ΔRGD (bottom). (III) Then, wild type (wt) HeLa cells (top) or wt fibroblasts (bottom) are seeded onto printed collagen I or fibronectin patterns for 60 min. Afterwards, weakly attached HeLa cells or fibroblasts are removed by gentle washing with PBS. (IV) merge of (I), (II) and (III). Scale bars, 20 μm. F Representative confocal microscopy images of fixed (I) wt HeLa cells adhering to collagen I patterns (pink) or (II) wt fibroblasts adhering to fibronectin patterns (pink) labeled for actin (yellow) and paxillin (blue). Scale bars, 15 µm. n = 10 independent experiments for each condition.

Back to article page