Lipid Dip-Pen Nanolithography for Model Bio-Membrane Systems

用于模型生物膜系统的脂质浸笔纳米光刻

• 批准号: 49885419
• 负责人: Professor Dr. Harald Fuchs
• 金额: --
• 依托单位: Physikalisches Institut
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/49885419?language=en
• 关键词: Lipid; Dip; Pen; Nanolithography; Model

The overall objective of this proposal is to merge top-down and bottom-up strategies to assemble nanoscale biomimetic lipid membranes relevant to immune transduction and cell adhesion. Specifically, we aim to develop tools for high throughput patterning of lipids on surfaces and to understand the fundamentals of lipid deposition and organization. Subsequently, we will use these capabilities to probe how geometrically-defined and chemically-tailored lipid structures affect cellular processes.There is growing evidence that the various properties of biological membranes strongly depend on both two- and three-dimensional heterogeneities (e.g. protein clustering, lipid rafts, and plasma membrane curvature). Furthermore, many cellular signaling processes respond to signaling agents at sub-cellular length scales. For studying spatially-resolved, supramolecular biological structure-function relationships, it is therefore increasingly important to interrogate these signaling events at the nanometer to micron length scales. We have previously developed Dip-Pen Nanolithography (DPN), a direct-write chemical deposition technique, capable of generating large-scale patterns with nanometer size features, and in the first three years of NSF-ICC funding, a toolkit for depositing lipids was developed. This lipid DPN (L-DPN) currently allows for the integration of two or more different membrane components on a surface with sub-cellular nanoscale spatial resolution and square centimeter per minute throughput. In addition, we have shown that L-DPN enables the investigation of cellular interactions with patterned surfaces that mimic biological structures.This proposal aims to exploit these biomimetic lipid patterns generated by L-DPN to examine two model cell signaling systems that are relevant to molecular biology and biochemistry: immunological mast cell activation by IgE; and cadherin-mediated epithelial cell adhesion. To fully enable these studies, additional multi-component lipid inks will be formulated and characterized according to deposition parameters and component distribution. The goals will be twofold: first, to answer specific fundamental questions related to cell-surface interactions; and second, to develop a new tool that will allow researchers to study these nanoscale interactions with high spatial control.

该提案的总体目的是合并自上而下和自下而上的策略，以组装与免疫转导和细胞粘附相关的纳米仿生脂质膜。具体而言，我们旨在开发用于表面上脂质的高吞吐量图案的工具，并了解脂质沉积和组织的基本原理。随后，我们将使用这些能力来探测几何定义和化学定量的脂质结构如何影响细胞过程。越来越多的证据表明，生物膜的各种特性在很大程度上依赖于二维和三维异质性（例如，蛋白质聚集，脂素，lipsma crafts和plasbrane membrane curvature）。此外，许多细胞信号传导过程在亚细胞长度尺度下响应信号传导剂。因此，为了研究空间分辨的超分子生物结构功能关系，因此将这些信号事件的纳米尺寸审问到微米长度尺度越来越重要。我们以前已经开发了一种直接处理化学沉积技术DIP-PEN纳米光刻（DPN），能够生成具有纳米尺寸特征的大型模式，并且在NSF-ICC资助的头三年中，开发了用于沉积脂质的工具包。该脂质DPN（L-DPN）当前允许在具有亚细胞纳米级空间分辨率和平方厘米每分钟吞吐量的表面上整合两个或更多不同的膜成分。此外，我们已经表明，L-DPN能够研究模仿生物结构的细胞相互作用。该建议旨在利用L-DPN生成的这些生物映射脂质模式，以检查两个模型细胞信号传导系统，这些模型细胞信号系统与分子生物学和生物学细胞化学：免疫学细胞：免疫学细胞激活by I ige by ige ige;和钙粘蛋白介导的上皮细胞粘附。为了充分启用这些研究，将根据沉积参数和组件分布制定和表征其他多组分脂质油墨。目标将是双重的：首先，回答与细胞表面相互作用有关的特定基本问题；其次，要开发一种新工具，该工具将允许研究人员研究这些纳米级相互作用，并具有高空间控制。