Microcontact Printing of Biomolecules on Various Polymeric Substrates: Limitations and Applicability for Fluorescence Microscopy and Subcellular Micropatterning Assays

Abstract

Polymeric materials play an emerging role in biosensing interfaces. Within this regard, polymers can serve as a superior surface for binding and printing of biomolecules. In this study, we characterized 11 different polymer foils [cyclic olefin polymer (COP), cyclic olefin copolymer (COC), polymethylmethacrylate (PMMA), DI-Acetate, Lumirror 4001, Melinex 506, Melinex ST 504, polyamide 6, polyethersulfone, polyether ether ketone, and polyimide] to test for the applicability for surface functionalization, biomolecule micropatterning, and fluorescence microscopy approaches. Pristine polymer foils were characterized via UV–vis spectroscopy. Functional groups were introduced by plasma activation and epoxysilane-coating. Polymer modification was evaluated by water contact angle measurement and X-ray photoelectron spectroscopy. Protein micropatterns were fabricated using microcontact printing. Functionalized substrates were characterized via fluorescence contrast measurements using epifluorescence and total internal reflection fluorescence microscopy. Results showed that all polymer substrates could be chemically modified with epoxide functional groups, as indicated by reduced water contact angles compared to untreated surfaces. However, transmission and refractive index measurements revealed differences in important optical parameters, which was further proved by fluorescence contrast measurements of printed biomolecules. COC, COP, and PMMA were identified as the most promising alternatives to commonly used glass coverslips, which also showed superior applicability in subcellular micropatterning experiments.

Cell Patterning Technology on Polymethyl Methacrylate through Controlled Physicochemical and Biochemical Functionalization

“…While some cells detached The possibility to avoid the blocking step allows the production and fabrication process of the platform to be sped up (from 3.5 to 1.5 h) and further enhances the potential for industrial-scale production of these microtechnologies. Furthermore, the incubation time required to generate the cell patterns has been reduced significantly from the previous reports on PMMA surfaces (from 4 h to 24 h down to 30-60 min [5,24]).…”

“…This greatly limits the commercialization potential of these microtechnologies. To create new commercial tools for the pharmaceutical and biomedical industries, it is desirable to extend the manufacturability of these novel microtechnologies to plastic substrates, which are less fragile and offer a wider range of optical properties [4,5].…”

“…Schmalenberg et al and Wang et al have reported successful applications of combined plasma treatment of PMMA surfaces and µCP of laminin for the adhesion and growth of Schwann cells and nerve cells, with high potential to function as nerve guides and promote nerve regeneration [23][24][25]. Recently, Hager et al studied subcellular patterning of different proteins through µCP on plasma-activated and epoxy-functionalized polymeric surfaces, including PMMA, and demonstrated that PMMA could be a feasible and flexible alternative to conventional cell culture materials, like glass, for µCP, presenting comparable optical properties [5]. Although there is growing interest in using these polymeric materials as substitutes for traditional cell culture materials in the development of novel cell monitoring microtechnologies, there has been limited exploration of the combination of physicochemical and biochemical modifications of PMMA surfaces to control cell adhesion on the typically cytophobic material.…”

Creating Supramolecular Functional Materials via Nucleobase Interaction: Investigating Molecular Recognition and the Fabrication of Surface Relief Gratings

“…8 Patterned films could be used directly in optical storage or solar energy transfer applications. 9,10 The formation of SRG relies on a cyclical isomerization and reorientation process of azobenzene-contained material. 5 That is, under specific light irradiation, azobenzene functionalities may undergo an E/Z conformational transition.…”

“…Photocontrollable surface structures of azo-containing material have shown exciting phenomena, such as controlled wettability, significant structural anisotropy, and light-harvesting ability . Patterned films could be used directly in optical storage or solar energy transfer applications. , …”