Hacking Morphogenesis: integrated, complex microsystems (ICS) for controlling embryonic pattern formation

该提案集中于1）分布式微尺度致动器、细胞内荧光传感器分子和反馈控制的集成系统的设计、制造和测试，该集成系统能够与发育中的斑马鱼胚胎组织化学地接口，（二）直接控制斑马鱼中胚层图案和胚胎发育。拟议的集成微系统将具有能够控制蛋白质扩散的化学致动器阵列组织中。该系统将通过将荧光蛋白序列拼接到报告细胞发育状态的两个基因（gsc-GFP和ntl-RFP）中来感测发育组织中的蛋白质产生水平。该系统将持续监测转录因子的状态，并调整蛋白质的分布剂量。PI将解决发育生物学中的一个基本开放问题：图灵/迈因哈特反应扩散模型是否适合描述斑马鱼中胚层图案？我们将探索斑马鱼胚胎如何产生复杂的分化模式，从而形成成熟的生物体。这项研究将影响发育的基础生物学，并为与发育中的生物体的相互作用提供根本性的新技术。虽然许多成熟的方法允许研究人员可视化发育中的组织，但用于控制发育中组织中的扩散环境的技术仍处于起步阶段。集成传感器/致动器组件阵列将使我们能够以亚细胞分辨率探测细胞反应。广泛的影响这一建议将促进不同实验室之间的交叉施肥。这次交流将包括在项目中工作的学生，并将包括棕色袋午餐和会谈的研究人员在每个机构的核心社区。密歇根纳米制造设施（MNF）将接待范德比尔特的研究人员，并向他们介绍先进的微/纳米制造方法。在范德比尔特大学，Boczko教授将接待微系统工程专业的学生，提供实验室轮换和斑马鱼胚胎生长、培养和观察方面的实验经验。

This proposal focuses on 1) the design, fabrication and testing of an integrated system of distributed microscale actuators, intracellular fluorescent sensor molecules and feedback control capable of chemically interfacing with developing Zebrafish embryonic tissue, 2) direct control of mesodermal patterning and embryonic development in Zebrafish.The proposed integrated microsystem will have an array of chemical actuators capable of controlling the diffusion of proteins into tissue. The system will sense the levels of protein production in the developing tissue by splicing fluorescent protein sequences into two genes that report on the developmental state of the cell (gsc-GFP and ntl-RFP). The system will continuously monitor the state of the transcription factors and adjust distributed dosing of proteins. The PIs will address a fundamental open question in developmental biology: is a Turing / Meinhardt reaction-diffusion model appropriate to describe mesodermal patterning in Zebrafish? We will explore how the Zebrafish embryo generates the complex patterns of differentiation that lead to a mature organism. Intellectual MeritThis research will impact the basic biology of development and provide fundamentally new technologies for interfacing with developing organisms. While a number of mature methods allow the researcher to visualize developing tissue, technologies for controlling the diffusible environment in developing tissue are in their infancy. Integrating arrays of sensor/actuator components will allow us to probe cell responses with sub-cellular resolution. Broad ImpactThis proposal will foster cross-fertilization between the different laboratories. This exchange will include the students working on the project and will include brown bag lunches and talks to the core community of researchers at each institution. The Michigan Nanofabrication Facility (MNF) will host Vanderbilt researchers and introduce them to advanced micro/nano-fabrication methods. At Vanderbilt University, Prof. Boczko will host microsystems engineering students, providing lab rotations and experimental experience in the growth, culture and observation of Zebrafish embryos.