Abstract
Lipid-based membranes play crucial roles in regulating the interface between cells and their external environment, the communication within cells, and cellular sensing. To study these important processes, various lipid-based artificial membrane models have been developed in recent years and, indeed, large-area arrays of supported lipid bilayers suit the needs of many of these studies remarkably well. Here, the direct-write scanning probe lithography technique called polymer pen lithography (PPL) was used as a tool for the creation of lipid micropatterns over large areas via polymer-stamp-mediated transfer of lipid-containing inks onto glass substrates. In order to better understand and control the lipid transfer in PPL, we conducted a systematic study of the influence of dwell time (i.e., duration of contact between tip and sample), humidity, and printing pressure on the outcome of PPL with phospholipids and discuss results in comparison to the more often studied dip-pen nanolithography with phospholipids. This is the first systematic study in phospholipid printing with PPL. Biocompatibility of the obtained substrates with up to two different ink compositions was demonstrated. The patterns are suitable to serve as a platform for mast cell activation experiments.
Highlights
Supported lipid bilayers (SLBs) have attracted much interest for decades as model membranes for studying processes in cell membranes and for biosensing applications [1], and—especially in recent years—as interfaces to living cells in biological and biomedical experiments [2]
While many experiments and applications can utilize uniform, large-area SLBs that are most often prepared by simple vesicle fusion approaches, more sophisticated setups can be realized by structured SLBs
While the general trends in regard to feature size result in regard to control parameters, such as dwell time, ambient humidity, and printing pressure applied to the stamp, hold true in comparison to Polymer pen lithography (PPL) printing with other materials, the dependence of dwell time and printing pressure was observed to be less pronounced in PPL with phospholipids
Summary
Supported lipid bilayers (SLBs) have attracted much interest for decades as model membranes for studying processes in cell membranes and for biosensing applications [1], and—especially in recent years—as interfaces to living cells in biological and biomedical experiments [2]. PPL can generate of lipid forming a patterningitprocess that allows for that arbitrary patterns by multipledifferent approaches types to the surface thatpatterns, can be precisely moved in lateral directions at the same time (insteadsizes havingofthe desired pattern gradient over mm scale distances, while maintaining subcellular the single features within the predefined on the stamp itself) and multiplexing [5,11,12,13]. Multiplexed lipid patterns of 1,2-dioleoyl-sn-glycero-3(as dwell time, environmental humidity, and printing pressure) would allow for a better and more phosphocholine (DOPC) admixed with different fluorophore-modified phospholipids on a micron rational approach generating scale in over large surface phospholipid areas (~cm2) printedmicro by PPLpatterns. Elucidating the influence of typical PPL process parameters (as dwell time, environmental the printing results are tested for feasibility in biological applications by a mast cell activation humidity, and printing pressure) would allow for a better and more rational approach in generating micro patterns. The printing results are tested for feasibility in biological applications by a mast cell activation demonstration experiment
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