Measuring single-cell water content non invasively and with high precision

无创高精度测量单细胞水分含量

• 批准号: 10711889
• 负责人: SCOTT R MANALIS
• 金额: $ 31.8万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711889
• 关键词: Acute; Area; Basic Science; Biochemistry; Biological; Biomedical Research; Cell Cycle; Cell Survival; Cell membrane; Cell physiology; Cells; Cellular biology; Complex; Crowding; Detection; Deuterium Oxide; Disease; Drug Exposure; Dryness; Exclusion; Exposure to; Fluorescence; Genetic Diseases; Growth; Health; Hour; Immune; Immunologic Surveillance; Immunomodulators; Liquid substance; Macrophage; Measurement; Measures; Methods; Mitosis; Molecular; Monitor; Mutate; Mutation; Normal Cell; Organoids; Outcome; Pharmaceutical Preparations; Phenotype; Process; Regulation; Research; Role; Sampling; System; Time; Water; biophysical properties; cell growth; cell water; cellular imaging; density; drug discovery; fluorescence imaging; in vivo; novel; particle; prevent; stem cell division; tool; tumor; virtual; water channel; water testing

SUMMARY Water is the medium in which biochemistry operates. The amount of water inside a cell defines the concentration of all biomolecules and the degree of molecular crowding. Consequently, water content is predicted to influence virtually all cell functions from immune surveillance to stem cell renewal in vivo. There are also severe genetic diseases caused by mutated water channels on cell membranes. Despite the fundamental role of cellular water content in cell physiology and diseases, there are no direct and non-invasive methods that measure how much water a single cell or a cluster of cells, such as an organoid, contains. This lack of water content measurements has prevented us from understanding cell physiology in normal and disease states, and from discovering drugs that can modulate cellular water content in diseases where water content is perturbed. Here, we propose to develop a new method that will directly, non-invasively and precisely measure the absolute and fractional (v/v) water content of a single cell or an organoid. This method will enable the long-term monitoring of the same cell or organoid during growth, differentiation or drug exposure. To achieve this, we will integrate total volume and dry volume measurements obtained using two independent approaches that we have previously developed. Our proposed method will enable water content measurements in complex and biomedically relevant samples, such as immune cells and organoids, with high throughput and in conjunction with detection of fluorescent markers. This method will enable novel basic and biomedical research that will increase our understanding of water content regulation in health and disease, and provide a new platform for drug discovery.

概括 水是生物化学运营的媒介。细胞内的水量定义了浓度 在所有生物分子和分子拥挤的程度中。因此，预计水含量会影响 几乎所有细胞功能从免疫监测到体内的干细胞更新。也有严重的遗传 由细胞膜上突变的水通道引起的疾病。尽管细胞水的基本作用 细胞生理和疾病中的含量，没有直接和非侵入性方法来衡量多少 为单个细胞或一个细胞（例如类器官）浇水。缺乏水含量测量 已经阻止了我们理解正常和疾病状态的细胞生理，也无法发现药物 这可以调节疾病的细胞水含量受到干扰。在这里，我们建议 开发一种新方法，该方法将直接，非侵入性，精确地测量绝对和分数（v/v） 单细胞或类器官的水含量。此方法将使对同一单元的长期监控 或在生长，分化或药物暴露期间的器官。为了实现这一目标，我们将整合总数和 使用我们先前已经开发的两种独立方法获得的干燥体积测量值。我们的 提出的方法将在复杂和生物医学相关样品中实现水含量测量，例如 作为免疫细胞和类器官，具有较高的吞吐量，并结合检测荧光标记。 这种方法将实现新颖的基本和生物医学研究，这将增加我们对水的理解 健康和疾病中的内容调节，并为药物发现提供了新的平台。