Microfluidics-Based Single-Cell Chemical Analysis of Cyanobacteria

单细胞分析是一项新兴技术，有望为生物学研究提供新的视角。由于单细胞分析是将细胞作为个体来观察的，因此它消除了集合平均并解决了样本群体内存在的异质性。这种异质性可以为重要的细胞过程提供有用的信息，特别是当大量细胞表现出与平均细胞行为的显著偏差时。本研究项目将在微流控平台上开发单细胞分析技术，研究单细胞藻胆酶体（PBS）的降解过程。通过单分子检测，可以直接从单细胞裂解液中计数非常低的荧光分子拷贝数，这是漂白蓝藻细胞的情况。这些工具将用于了解单细胞水平PBS降解的生化机制。更广泛的影响这个项目将对许多不同的研究领域产生重大影响。首先，从本研究中获得的生物学见解将为观察生态多样性、微生物学及其相互关系提供新的途径。其次，单细胞技术将向细胞培养、细胞分离和操作以及细胞内内容物（包括蛋白质和核酸）分析的整合方向发展。通过这种综合方法，应该可以从单个细胞以及简单的表型变化中获得准确的生化信息。这种策略可以扩展到其他类型的细胞，并为研究生物异质性提供了一个新的范例。第三，这个项目本质上是跨学科的，涉及化学分析和生物学结果。这种结合应该会在许多领域之间引发协同对话，包括生态学、种群生物学和生物物理化学，从而为未来的研究带来更富有成效的方向和动力。第四，在本科和研究生阶段，将有一个强有力的教学和培训组成部分，以及每年的多学科研讨会，向参与者介绍单细胞技术在生物学中的潜在用途。

Single-cell analysis is an emerging technology that promises to provide new perspectives on biology. Because single-cell analysis observes cells as individuals, it eliminates ensemble averaging and resolves the heterogeneity present within a sample population. This heterogeneity can provide useful information with regard to important cellular processes, especially if a significant number of cells display a marked deviation from that of average-cell behavior. This research project will be to develop single-cell analysis technology on a microfluidic platform to study the phycobilisome (PBS) degradation process of a unicellular cyanobacterium Synechococcus. With the single-molecule detection, very low copy numbers of fluorescent molecules can be directly counted from a single-cell lysate, which is the case for bleached cyanobacteria cells. These tools will be used to understand the biochemical mechanism of PBS degradation at the single-cell level.Broader ImpactsThis project will have a significant impact on many different areas of research. First, the biological insight gained from this research will illuminate a new way of viewing ecological diversity, microbiology, and the interrelationship between them. Second, single-cell technology will be advanced toward the integration of cell culture, isolation and manipulation of cells, and the analysis of intracellular contents (both proteins and nucleic acids). With this integrative approach, it should be possible to obtain accurate biochemical information from individual cells as well as simple phenotypic changes. This strategy can be extended to other types of cells and should provide a new paradigm for studying biological heterogeneity. Third, this project is interdisciplinary in essence, involving chemical analysis with biological outcomes. This combination should induce a synergistic conversation between many fields, including ecology, population biology, and biophysical chemistry, resulting in more productive directions and motivations for future research. Fourth, there will be a strong component of teaching and training both at the undergraduate and graduate level as well as annual multidisciplinary workshops to introduce participants to the potential uses of single-cell technology in biology.