Reusable, reversibly sealable parylene membranes for cell and protein patterning

Abstract

The patterned deposition of cells and biomolecules on surfaces is a potentially useful tool for in vitro diagnostics, high-throughput screening, and tissue engineering. Here, we describe an inexpensive and potentially widely applicable micropatterning technique that uses reversible sealing of microfabricated parylene-C stencils on surfaces to enable surface patterning. Using these stencils it is possible to generate micropatterns and copatterns of proteins and cells, including NIH-3T3 fibroblasts, hepatocytes and embryonic stem cells. After patterning, the stencils can be removed from the surface, plasma treated to remove adsorbed proteins, and reused. A variety of hydrophobic surfaces including PDMS, polystyrene and acrylated glass were patterned using this approach. Furthermore, we demonstrated the reusability and mechanical integrity of the parylene membrane for at least 10 consecutive patterning processes. These parylene-C stencils are potentially scalable commercially and easily accessible for many biological and biomedical applications.

Selective hydrophilic modification of Parylene C films: a new approach to cell micro-patterning for synthetic biology applications

“…Microfabricated polymer stencils is another method to engineer the cellular microenvironment by serving as selective physical barriers that are peeled-off after seeding. Due to its high Young's modulus, Parylene C has been widely used to pattern proteins [11,12] and cells [12]. Nonetheless, this technique involves a very delicate procedure to peel-off the Parylene stencil intact, which often results in partial destruction of the polymer membrane and residual pieces on the supporting surface especially when thin (~ 1 µm) stencils are used.…”

Block-Cell-Printing for live single-cell printing

“…Several approaches have been designed for this purpose, e.g., inkjet cell printing (3)(4)(5)(6), surface engineering (7)(8)(9)(10)(11)(12)(13)(14)(15), and physical constraints (16)(17)(18)(19)(20)(21)(22)(23). However, finding a method that completely satisfies the above requirements remains a challenge.…”

Fabrication of multi-layer polymeric micro-sieve having narrow slot pores with conventional ultraviolet-lithography and micro-fabrication techniques

“…For instance, it can be used as a stencil for micro-patterning of cells, bio-molecules and metallic films 27 or even can be employed for separation of blood cells (red blood cells (RBCs) and white blood cells (WBCs)) from whole blood for clinical assays. 28 …”