Precision Multi-site Cellular Dosing using a Membrane-Based Laminar-Flow Device

描述(由申请人提供)：拟议的第一阶段SBIR项目旨在展示开发一种负担得起的商业仪器的可行性，该仪器可实时在多个坐标下为细胞培养提供层流流体界面。该仪器将满足日益增长的、尚未得到满足的对一种方法的需求，这种方法允许使用廉价的材料和设备对化学剂量和细胞之间的相互作用进行卓越的时空控制。时空控制对于检测化学剂量与细胞之间的相互作用至关重要，特别是在专注于单个细胞内的极化反应的研究中，例如定向趋化。Minotaur Technologies的方法是基于激光在膜内快速创建孔，作为细胞生长的支撑，并分隔两个流动室-其中一个是细胞培养室，另一个是配药室。这种使用激光诱导的介电击穿作为加工表面孔的手段允许从高压配料腔精确定向的层流流入电池环境，从而仅向某些选定的电池或电池区域添加所需的化学物质。在第一阶段项目中，Minotaur将展示剂量区域分辨率降至2微米的细胞剂量的可行性，并展示该技术在重要的神经科学应用中的好处。尽管我们在分子水平上了解了细胞内部的许多情况，但对细胞作为一个整体的功能的理解却远远落后。活细胞的研究常常因为缺乏适当的技术而受到阻碍。科学进步不仅依赖于新思想，还依赖于使观念转变成为可能的技术进步。在活细胞中加入化学剂量的试剂，选择性地以受控的方式改变细胞功能，为研究细胞的分化、生长和死亡提供了重要的手段。开发的仪器将适用于神经科学和其他领域的广泛细胞生物学问题，包括趋化性、干细胞分化和对新出现的治疗方法的细胞反应的体外测试。与公共卫生的相关性：细胞生物体的一个基本组成部分是细胞感知和响应环境的能力。阐明细胞对外界刺激的反应机制以及细胞间信号通路如何协调细胞活动是理解细胞如何工作的关键。通过使用拟议的仪器，可以更好地了解健康和疾病状态下的细胞反应，可能会导致开发更有效的药物和诊断方法。

DESCRIPTION (provided by applicant): The proposed Phase I SBIR project is to show the feasibility of developing an affordable commercial instrument that provides laminar-flow fluidic interfaces to cell cultures at multiple coordinates in real time. The instrument will satisfy a growing, unmet need for a method that allows superior spatio-temporal control over the interactions between chemical dosants and cells using inexpensive materials and equipment. Spatio-temporal control is critical to examining interactions between chemical dosants and cells, especially in studies focused on polarized responses within individual cells such as directional chemotaxis. Minotaur Technologies' approach is based on the rapid laser-mediated creation of apertures within a membrane that serves as a support for cell growth and also separates two flow chambers - one of which is a cell medium chamber, and the other, a dosing chamber. This use of laser-induced dielectric breakdown as a means to machine surface pores allows for a precisely-directed laminar flow stream from the higher pressure dosing chamber into the cell environment such that only certain selected cells or cell regions are dosed with the desired chemical species. In the Phase I project Minotaur will demonstrate the feasibility of dosing cells with a dosant region resolution down to 2 micrometers and show the benefits of the technology for important neuroscience applications. Although we understand much of what goes on inside cells at a molecular level, understanding of how cells function as a whole lags far behind. Frequently, the study of living cells is hampered by lack of appropriate technologies. Scientific advances depend not only on new ideas, but also on technological advances that make conceptual shifts possible. Chemical dosing of live cells with agents that selectively alter cellular function in a controlled fashion provides a vital means to study cellular differentiation, growth, and death. The instrument developed will be applicable to a broad range of cell biology problems in neuroscience and beyond, including chemotaxis, stem-cell differentiation and in vitro testing of cellular response to emerging treatments. Public Health Relevance: A fundamental component of cellular organisms is the ability of cells to sense and respond to their environment. Elucidating the mechanisms of cellular responses to external stimuli and how cell-to-cell signaling pathways coordinate cellular activities is key to understanding how cells work. The improved understanding of cellular responses in both healthy and diseased states that may be gained from use of the proposed instrument could lead to development of more effective medicines and methods of diagnosis.