Currently, cell patterning and isolation methods often utilize microfluidic systems, in which fluid forces are used to manipulate and transport cells. Inkjet-based cell "printing" and deposition methods have been shown to be effective at sorting and patterning cells at the bulk and single-cell level, but are generally low-throughput and can cause cellular stress responses. Additionally, although Trap-based methods have high throughput, but they may discriminate between specific cell morphologies or sizes associated with human disease. The microfluidic trap environment also makes it difficult to transport individual cells into isolated microenvironments for further experiments. Another disadvantage of all microfluidic patterning is that they subject cells to shear stress that can affect cell activity, function, and gene expression.

In contrast to these methods, cell patterning stencil can facilitate high-throughput single-cell analysis. Because this method does not require surface modification and relies only on physical barriers, cell size can be displayed indiscriminately as long as the stencil through-holes are large enough. In addition, the stencil maintains the normal morphology of neurons, providing a space between the glass substrate and the stencil membrane for cell axonal growth.

Fig 1. Generation of PDMS stencil for micropatterning.

Our Cell Patterning Stencil

The main strategy of Creative Bioarray cell patterning stencil is a physical barrier based on PDMS, which is not only soft, transparent, and bendable, but also works on various surfaces, even curved substrates. When we use this method, the stencil is immobilized on the substrate surface prior to cell seeding. In the through-pore region, the diameter of the pores is slightly larger than or approximately equal to the cell size, and cells can settle through the pores and pattern on accessible areas of the surface at the single-cell level. In other areas that serve as physical barriers, cells are blocked by the template film. When cells adhere to the surface, a high-throughput single-cell array can be formed after removing the stencil, which will greatly facilitate your high-throughput single-cell analysis.

Additionally, we can configure single, double, triple or multi-cell configuration within each space by varying pattern size, cell suspension density, and droplet volume depending on your experimental purpose.

Our Advantages

• Does not put any additional stress on cells and does not reduce cell viability
• Greatly improves the efficiency of seeding cells to defined locations on substrates
• Making high-throughput single-cell analysis easier
• Simultaneous acquisition of cellular response information at the population, single cell and subcellular levels
• Spatial organization that allows probing of differentiation fate
• Ideal for studying multicellular tissue patterning during different developmental processes (e.g., neuroepithelial folding or endoderm patterns)

Related Services

• Single-cell Patterning
• Cell Patterning for Controlling Cell-cell Interactions
• Cell Patterning for Regulating Cell Differentiation

References:

1. Sahni G, et al. Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates. J Vis Exp. 2016, (112):54097.
2. Messner, J.J, et al. Laser-fabricated cell patterning stencil for single cell analysis. BMC Biotechnol. 2017,17(1).