权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Sorting Sounds - A high-throughput microfluidics screening platform for the development of genetically encoded labels for Optoacoustic imaging

Sorting Sounds - 高通量微流体筛选平台，用于开发用于光声成像的基因编码标签

基本信息

批准号：
323341449
负责人：
Dr. Andre Stiel, Ph.D.
金额：
--
依托单位：
Institut für Biologische und Medizinische Bildgebung (IBMI)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2016
资助国家：
德国
起止时间：
2015-12-31 至 2021-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/323341449?language=en
关键词：
Sorting Sounds throughput microfluidics screening

项目摘要

Optoacoustic (OA) or Photoacoustic imaging is an emerging imaging modality providing unsurpassed penetration depth at high resolutions. This provides for more comprehensive time resolved 3D in vivo imaging at depths far beyond the reach of optical methods. However, with the application of OA in life sciences being relatively new the range of labels and molecular tools is limited. Thus far only a small number of dedicated labels has been put forward with an even smaller number being genetically encoded - a prerequisite for targetable in vivo imaging. From the molecular perspective a wide range of labels known from fluorescence imaging - especially green fluorescent protein (GFP) like labels - could be utilized for OA. However, while fluorescence imaging aims for a high molecular brightness (i.e. quantum yield) the concept of OA requires a predominantly radiation-less deexcitation pathway. This fundamental difference makes an immediate application of the rich variety of fluorescence imaging tools in OA problematic. Furthermore, near-infrared labels are required to access the full advantage of the higher penetration depth intrinsic to OA imaging. Consequently, a new range of molecular labels has to be designed for OA to fully empower this new imaging modality. Next to the design principles a fundamental prerequisite to protein based label engineering is the ability to screen for the desired properties of the label. In the case of the design of genetically encoded fluorescence labels screening could rely on the detection of fluorescence signals. However, new screening strategies have to be applied for OA-label engineering. The research outlined in this proposal includes design of a microfluidics based high-throughput OA cytometry device as well as proof-of-concept directed-evolution and screening for OA-optimized labels. In preliminary work we could already demonstrate the feasibility of faithfully detecting OA signals in single cells in a microfluidics configuration and outlined the technical prerequisites. Furthermore, we tested several red GFP-type chromo-proteins in OA to identify candidates most suitable as starting points for library generation.

光声（OA）或光声成像是一种新兴的成像方式，提供无与伦比的穿透深度在高分辨率。这提供了在远远超出光学方法所能达到的深度处的更全面的时间分辨3D体内成像。然而，随着OA在生命科学中的应用相对较新，标签和分子工具的范围受到限制。到目前为止，只有少量的专用标签已被提出，甚至更少的数字是遗传编码-一个先决条件，在体内成像的目标。从分子的角度来看，从荧光成像已知的广泛的标记-特别是绿色荧光蛋白（GFP）样标记-可以用于OA。然而，虽然荧光成像的目标是高分子亮度（即量子产率），但OA的概念需要一个主要的无辐射去激发途径。这种根本的差异使得在OA中立即应用丰富多样的荧光成像工具成为问题。此外，需要近红外标签来获得OA成像固有的更高穿透深度的全部优势。因此，必须为OA设计一系列新的分子标记，以充分增强这种新的成像方式。除了设计原则之外，基于蛋白质的标签工程的基本先决条件是筛选标签的所需性质的能力。在设计遗传编码的荧光标记的情况下，筛选可以依赖于荧光信号的检测。然而，新的筛选策略必须应用于OA标记工程。该提案中概述的研究包括基于微流体的高通量OA细胞仪设备的设计以及OA优化标记的概念验证和定向进化和筛选。在初步工作中，我们已经证明了在微流体配置中忠实检测单细胞中OA信号的可行性，并概述了技术先决条件。此外，我们测试了OA中的几种红色GFP型色素蛋白，以鉴定最适合作为文库生成起点的候选物。