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

体外平台是回答有关生物学和疾病的基本问题的关键工具。制造复杂细胞培养系统的策略之一是微图案化，它能够在体外控制细胞和组织结构，以探索和剖析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不同领域产生的功能和行为配置。微图案化的两种主要技术--微合同印刷和深紫外微图案化，需要复杂、多步骤、人工操作，孵化时间长，应用和重现性有限，不适合高通量分析，限制了其应用和开发。作为其生物材料和微流体服务的一部分，Making Lab希望利用分子等离子体图案化的最新进展，提出一种尖端的全自动化设备，其快速实现、操作简单和通用性将立即有利于Crick冷大气分子等离子体的研究。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嫁接各种化学(抗体、多肽、蛋白质、环氧体、等)，在室温下以一步、无溶剂、可扩展的大气工艺直接沉积到任何衬底上。此外，这一工艺可以适用于生物学中使用的任何类型的载体，包括S具有挑战性的用于小容量或高通量实验的载体。结合微图案掩膜(即具有负特征的掩膜，让等离子体和分子压印在载体表面)、具有同轴生物分子沉积的冷大气等离子体以及计算机数控(NC)制造系统，它允许编程和自动化表面处理。在实践中，使用惰性气体流来产生通过载体表面的图案投影的等离子体，同时同轴分布包含气雾剂形式的分子的溶液以结合到表面的等离子体上。涂布程序是在机器软件上编程的，可以适应任何类型的表面和适合A4表面的碟子。放置在等离子顶部的定制适配器允许使用多个头，以允许在整个表面、仅部件或图案上进行涂层。建议为该仪器开发一种定制写入等离子体头以及将其应用于生物学研究的相关协议，这是一个独特的机会，可以显著增加该仪器对新项目开发的影响。这项技术将立即惠及生物工程、发育生物学、细胞生物学、感染和免疫、基因调控、神经科学、纳米制造和生物传感等领域的实验室。例如，它将有助于更好地了解人类巨噬细胞和结核分枝杆菌(MTB)在早期感染状态下的动态相互作用，或描述血管拓扑背后的动态，有助于找到更准确地对神经系统进行建模的方法，了解运动神经元疾病(MND)等条件如何损害它，以及自动检测3D细胞培养阵列，以便将其用于药物发现。

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.