Micropatterning of adhesion proteins at the subcellular level

Create micropatterns of adhesion proteins with a resolution of 1.2 μm thanks to PRIMO.

Micro-dots-micropatterns
Epifluorescence microscopy image of cells expressing a transmembrane protein fused to GFP accumulated on micrometric dots.

Single cell micropatterning

Isolating single cells under reproducible conditions is fundamental for in vitro cell assays, with applications such as mechanobiology, the study of intracellular mechanisms or the screening of molecules.

Micropatterning of adhesion proteins at a cellular and subcellular scale with PRIMO system addresses this issue precisely with a very high flexibility of usable patterns and proteins.

fibronectin fibrinogen patterns
Embryonic fibroblasts from vimentin knockout mice on fibronectin + fibrinogen-A647 (blue) patterns of different shapes, actin labeled with phalloidin-A555 (red) and focal adhesions revealed via Anti-Paxillin antibodies + secondary antibodies coupled to A488 (green). Courtesy of A.J. Jimenez and B. Vianay, Physics of cytoskeleton & Morphogenesis lab.
micropattern of fibronectin
Fibroblast adhering on a micropattern of fibronectin, actin labeled in red.
Fibroblasts plated on fibronectin micropatterns
Fibroblasts plated on fibronectin micropatterns (after 2 weeks).

Single cell patterning for neurosciences.

1.Astrocyte-based assays conducted first without micropatterning:

fibronectin-coating
Astrocytes plated on a uniform fibronectin coating show heterogeneity in shapes. Courtesy of C. Delépine. Delépine C. et al., Hum. Mol. Genet. 2016.

The microtubules were tagged with GFP, but their growth direction was heterogeneous and couldn’t be measured. Courtesy of C. Delépine. Delépine C. et al., Hum. Mol. Genet. 2016.

2.Astrocyte-based assays conducted with PRIMO micropatterning:

Astrocytes-fibronectin-patterning
Astrocytes plated on fibronectin lines. Courtesy of C. Delépine. Delépine C. et al., Hum. Mol. Genet. 2016.

The microtubules were tagged with GFP and then shown the same orientation, which allowed to measure microtubule growth rates. Courtesy of C. Delépine. Delépine C. et al., Hum. Mol. Genet. 2016.

Patterning of cell populations

On a larger scale, PRIMO also makes it possible to control complex cellular arrangements.

fibronectin micropatterns
Mouse teratocarcinoma cells plated on fibronectin micropatterns.
grid of fibronectin micropattern
Epithelial cells on fibronectin stained for actin (green), nuclei (blue) and focal adhesions (red), on a grid of fibronectin micropattern.

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