Super plan fluor 20 elwd lens

Micropatterned adhesive surfaces were generated using the PRIMO optical module (Alvéole, France) controlled by the Leonardo plugin (V3.3, Alveole) mounted on a Nikon TI-E inverted microscope (Nikon Instruments) equipped with a Super Plan FLuor ×20 ELWD lens (Nikon) lens and a DMD-based UV (375 nm). To generate circular and square micropatterns, both 100 µm diameter, were projected onto plasma-cleaned (Corona Treater, ETP), PLLgPEG-passivated (0.1 mg/ml PLL-g-PEG (PLL (20)-g [3.5]- PEG (2), SuSoS) 35 mm glass-bottom dishes (Ibidi). Patterned areas were then washed multiple times with PBS and conjugated with a uniform coating of 10 µg/ml fibronectin and 38.75 µg/ml collagen for 1 h at +37 °C. The substrates were then washed with PBS, prior to seeding 100 K of mouse keratinocytes onto each 35 mm dish. Keratinocyte monolayers were allowed to proliferate on the patterns for 24 h, at which time they were subjected to AFM or fixed and processed for immunofluorescence and quantification analyses. For the analysis of YAP localization (Fig. 3d, e), only micropatterns with similar cell densities were compared.

Photopatterning of soft PDMS gels was performed using the PRIMO optical module^{59 (link)} (Alvéole) controlled by the Leonardo plugin (Alvéole) mounted on a Nikon inverted microscope (Nikon Instruments) equipped with a Super Plan Fluor ×20 ELWD lens (Nikon) and a DMD-based UV (375 nm). Before starting, the liquid on each sample was carefully aspirated (without letting the sample dry) and the sample was covered with PLPP photoactivator (Alvéole). The desired patterns for photoillumination were created using Inkscape (Inkscape Project) and loaded into Leonardo. The UV dose of all samples was set to 900 mJ/mm². After photopatterning, samples were rinsed 4 times with phosphate-buffered saline (PBS, Sigma-Aldrich), incubated for 5 min with a 0.02% fibronectin (F0895, Sigma-Aldrich) solution in PBS and rinsed thoroughly 5 times with PBS. For timelapse imaging experiments, filtered (220 nm filter) Alexa Fluor 647 conjugated fibrinogen was added to the fibronectin solution to allow visualization of the micropattern. Samples were stored at 4 °C until use (less than 48 h).