Automated Direct Micropatterning Platform for bespoke in vitro cell microenvironment using Molecular Plasma

体外平台是回答生物学和疾病基本问题的关键工具。产生复杂细胞培养系统的策略之一是微模式，能够在体外控制细胞和组织结构，以探索和解剖细胞和组织结构之间的关系，以及在疾病建模、免疫学或神经生物学等各个领域产生的功能和行为配置。用于微图案的两种主要技术，微收缩印刷和深紫外微图案，需要复杂，多步骤，手动操作，长时间的培养，并且在应用和重现性方面受到限制，因此不适合高通量分析，限制了它们的应用和发展。作为其生物材料和微流体服务的一部分，制造实验室希望利用分子等离子体的最新进展来提出一种尖端的全自动设备，其快速实施，操作简单和多功能性将立即有利于CrickCold大气研究分子等离子体是一种有趣的技术，它使用等离子体作为载体来移植各种化学物质(抗体，肽，蛋白质，环氧树脂，丙烯酸，等)在单步、无溶剂、可扩展的室温常压工艺中直接涂在任何基材上。此外，该过程可以适用于生物学中使用的任何类型的支持，包括用于低体积或高通量实验的具有挑战性的支持。结合微图案掩模（即具有负面特征的掩模，让等离子体和分子在支架表面上留下印记），具有同轴生物分子沉积的冷大气等离子体和计算机数控（CNC）制造系统，它可以实现表面处理的编程和自动化。在实践中，惰性气体流被用来产生通过支撑表面图案投射的等离子体，而含有气溶胶形式分子的溶液被同轴分布以与表面上的等离子体结合。涂层顺序在机器的软件上编程，可以适应任何类型的表面和盘子适合于A4表面。放置在等离子体顶部的定制适配器允许使用多个头，以允许整个表面，部分或图案的涂层。该仪器的定制书写等离子体头的建议开发以及将其应用于生物研究的相关协议代表了一个独特的机会，可以显着增加该仪器对新项目开发的影响。最终，该设备可以解决广泛的领域，如细胞生长，细胞迁移，器官，微结构衬底和微流体的微加工。这项技术将立即使生物工程、发育生物学、细胞生物学、感染和免疫、基因调控、神经科学、纳米制造、生物传感等领域的实验室受益。例如，它将有助于更好地理解人类巨噬细胞与结核分枝杆菌（Mtb）在早期感染状态下的动态相互作用，或描述血管拓扑结构背后的动态，有助于找到更准确地模拟神经系统的方法，并了解运动神经元疾病（MND）等疾病如何损害神经系统，以及自动测定3D细胞培养阵列，使其能够用于药物发现。

In vitro platforms are key tools to answer fundamental questions about biology and diseases. One of the strategies to produce a complex cell culture system is micropatterning, enabling control over cell and tissue architecture in vitro to explore and dissect the relationship between cell and tissue architecture and resulting function and behaviour configurations in various fields such as disease modelling, immunology, or neurobiology. The two main techniques used for micropatterning, micro contract printing and deep UV micropatterning, require complex, multistep, manual operations, lengthy incubation time, and are limited in application and reproducibility therefore not suitable for high throughput assays and limited in their applications and development.As part of its Biomaterial and Microfluidics service, the Making Lab wishes to take advantage of the recent advances in molecular plasma for patterning to propose a cutting-edge fully automated equipment, whose rapid implementation, simplicity of operation and versatility will immediately benefit research at the CrickCold atmospheric Molecular plasma is an interesting technology that use the plasma as a vector to graft various chemistry (antibodies, peptides, proteins, epoxy, acrylic, etc) directly onto any substrate in a single-step, solvent-free, scalable atmospheric process at room temperature. Moreover, this process can be adapted to any type of support used in biology including s challenging ones used for low volumes or high throughput experiments.Combining micropatterning masks (i.e. mask with negative features letting the plasma and molecules imprint them on the support's surface), a cold atmospheric plasma with coaxial biomolecule deposition, and a computer numerical control (CNC) manufacturing system, it allows programming and automation of surface treatment.In practice, a stream of inert gas is used to create a plasma projected through a pattern on the support's surface while a solution containing the molecule in aerosol form is coaxially distributed to bind to the plasma on the surface. The coating sequence is programmed on the machine's software and can be adapted to any type of surface and dish fitting into an A4 surface. A custom adapter placed on top of the plasma allows the use of multiple heads to allow the coating of the entire surface, parts only or patterns. The proposed development of a custom writing plasma head for the instrument and the associated protocols to apply it to biological research represents a unique opportunity to significantly increase the impact that this instrument could have on the development of new projectUltimately this equipment allows to address a wide range of areas such as cell growth, cell migration, organdies, microfabrication of microstructures substrate and microfluidics. This technology will immediately benefit laboratories working in the areas of bioengineering, developmental biology, cell biology, infection and immunity, gene regulation, neuroscience, nanofabrication, biosensing. For example, it will help better understand the dynamic interactions between human macrophages and Mycobacterium tuberculosis (Mtb) during early infection state, or describe the dynamic behind the vascular topology, contribute to finding ways to model the nervous system more accurately and see how conditions like motor neurone disease (MND) damage it and to automatically assay arrays of 3D cell cultures to enable their use in drug discovery.