Technology development for in situ imaging of microbial cultures

目前用于估计细菌培养物生长的标准，光密度（OD）测量，不再提供学术研究，工业生物技术应用和抗菌药物耐药性药理学工作所需的足够信息。事实上，所有这些领域都越来越需要关于细胞形态的准确信息，细胞形态现在被认为是对产品产量有负面影响的宿主细胞生理变化的早期指标。此外，特定的形状变化指向导致它们的潜在分子机制，从而能够更有效地对产品开发途径进行故障排除。例如，在大肠杆菌中生产重组蛋白会影响细胞的细胞内压力，迫使细胞在发酵过程中无意中泄漏产物和其他细胞内容物。细胞压力变化的早期指示是细胞大小的增加和随后导致细胞起泡的细胞形状的特征性变化，如果早期检测到，则可以解释。此外，在生产过程中对细胞进行成像可以改善对产物积累的分析（例如早期检测包涵体），同时提供形态信息，从而有可能揭示导致其产生的分子途径。为了克服这一挑战，作为该项目的一部分，您将开发细胞计数器/成像仪设备。全球细胞计数和高通量成像市场目前主要集中在哺乳动物细胞上，而在这里，我们将重点关注微生物细胞成像。微生物，特别是细菌细胞的成像（和通过成像计数）任务是不平凡的，因为它们的尺寸小，尺寸变化大（高达100倍），以及细菌可以游泳的事实。您将在由物理学家，工程师，生物技术专家和微生物学家组成的跨学科团队中工作，以在该项目中取得成功，并将学习他们在定制显微镜和微生物学方面的广泛专业知识。由于该项目具有商业潜力，日常工作的细节是保密的。请直接给Teuta发电子邮件，谈谈项目时间轴和所涉及的具体技术（teuta. ed.ac.uk）

The current standard for estimating the growth of bacterial cultures, optical density (OD) measurements, no longer provides sufficient information needed for academic research, industrial biotechnology applications, and pharmacological work on antimicrobial resistance. As a matter of fact, all these sectors increasingly require accurate information on cell morphology, which is now recognized as an early indicator of host-cell physiological changes that have a negative effect on the product yield. Additionally, specific shape changes point to the underlying molecular mechanism that causes them, enabling more efficient troubleshooting of product development pathways. For example, production of recombinant proteins in Escherichia coli affects cells' intracellular pressure, forcing the cells to inadvertently leak the product and other cellular content during fermentation. An early indication of cellular pressure changes is increase in the cells size and a characteristic change in cell shape that subsequently leads to cell blebbing, which, if detected early, can be accounted for. Furthermore, imaging cells during production can enable improved analysis of product accumulation (such as early detection of inclusion bodies) simultaneously with morphological information, which offers the potential to reveal the molecular pathway that causes it. To overcome this challenge, as part of this project you will develop a cell counter/imager device. The global cell counting and high-throughput imaging market is currently focused mainly on mammalian cells, whereas here we will focus on imaging microbial cells. The task of imaging (and counting via imaging) of microbial, and in particular bacterial cells, is non-trivial due to their small size, extensive size changes (up to 100 times), as well as the fact that bacteria can swim. You will work in an interdisciplinary team composed of physicists, engineers, biotechnologists and microbiologists to succeed in this project, and will learn from their extensive expertise in customized microscopy and microbiology. Since this project has commercial potential, details of day to day work are confidential. Please email Teuta directly to speak about the project timeline and specific techniques involved (teuta.pilizota@ed.ac.uk)