Nanostructured substrates for cell assays.

描述(由申请人提供)：该第一阶段SBIR的长期目标是开发优化的组件(纳米结构底物)，用于创建一种新型的基于液晶的快速细胞计数、增殖和功能分析。需要劳动密集度较低、消耗更少的细胞和试剂、能够在更短的时间内完成、允许将趋化作用与趋化作用分离、提供对浓度梯度传递的空间和时间方面的更好控制以及能够适应高通量格式的新技术。鸭嘴兽技术(Tm)可以作为此类分析的基础，以加速基础细胞研究、药物发现和抗癌药物的评估。纳米结构底物与液晶的结合不需要标记或额外的试剂来报告细胞的存在。它将为许多不同的细胞分析提供一个统一的平台。在这个方案中，我们将通过三种方法来制备具有纳米级形貌的衬底：纳米模塑、聚合物材料磨损和倾斜沉积金薄膜。我们将比较评估这些底物的能力：在细胞介质中存在蛋白质非特异性吸附的情况下排列液晶，支持上皮细胞、成纤维细胞和血管内皮细胞的细胞功能，报告细胞数量，并在六个月的保存期内保持功能稳定性。在完成这些研究后，我们将优化细胞计数和功能评估新技术的核心组成部分。

DESCRIPTION (provided by applicant): The long-term goal of this Phase I SBIR is to develop optimized components (nanostructured substrates) for use in the creation of a novel class of rapid, liquid crystal based cellular assays of enumeration, proliferation and function. There is a need for new technologies that are less labor intensive, consume fewer cells and reagents, can be performed in less time, allow for separation of chemotaxis from chemokinesis, provide better control over spatial and temporal aspects of the delivery of concentration gradients and are amenable to high throughput formats. Platypus Technology (tm) can be the foundation for such assays to accelerate basic cell research, drug discovery, and evaluation of anti-cancer drugs. The combination of nanostructured substrates with liquid crystals requires no labels or additional reagents to report the presence of cells. It will provide a unified platform for many diverse cellular assays. In this proposal, we will fabricate substrates with nanoscale topography by three methods: nanoscale molding, abrasion of polymeric materials and oblique deposition of thin films of gold. We will comparatively evaluate the ability of these substrates to: align liquid crystals in the presence of non-specific adsorption of proteins in cell media, support cell functions of epithelial, fibroblast and vascular endothelial cells, report cell numbers, and retain functional stability during a six-month storage period. At the completion of these studies we will have optimized the core component of a new technology for enumeration and functional evaluation of cells.