单细胞器官提取与细胞索引.
Trinh Lam1, Alison Su1, Ana E Gomez Martinez1
1Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.
bioRxiv : the preprint server for biology
|January 7, 2025
概括
使用一种新型的微流体装置,VacTrap从哺乳动物细胞中分离和空间索引单个核. 这一突破使得核的精确映射到它们的起源细胞,用于先进的多组学研究.
科学领域:
- 细胞生物学 细胞生物学
- 生物技术是生物技术.
- 微流体学 微流体学
背景情况:
- 大量有机体分成方法缺乏单细胞分辨率.
- 现有的微流体技术无法将分离器官与它们的起源细胞联系起来.
- 单细胞多组学需要精确地映射细胞组件回到它们的源细胞.
研究的目的:
- 开发一种高通量微流体装置,用于隔离和空间索引单个核.
- 为了使单个细胞核与它们的原始哺乳动物细胞进行映射.
- 为推进单细胞多组应用提供一个工具.
主要方法:
- 开发VacTrap,这是一个三层微流体装置,使用带有"陷门" (BAC-gel) 的Bis-gel微和真空分流器.
- 细胞溶解使用DDF释放细胞核,随后是细胞质蛋白的电泳分离.
- 通过真空辅助,同步的核从Bis-gel转移到PDMS微,并溶解陷门.
主要成果:
- 在80%的256微波阵列中,VacTrap实现了98%的同步核转移效率.
- 与被动方法 (<1%) 相比,核转移效率超过1%.
- 形态分析证实,在VacTrap过程中,有机细胞的完整性得到了保护.
结论:
- VacTrap提供了一个强大的,高吞吐量解决方案,用于单个核的空间索引.
- 该装置能够精确地将核与其起源细胞进行映射,这对于多组学至关重要.
- 通过保留细胞背景,VacTrap推进了单细胞分析领域.
相关概念视频
Subcellular Fractionation
6.8K
The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...
Differential Centrifugation
Differential centrifugation is...
6.8K
Overview Of Cell Separation And Isolation
5.5K
Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
5.5K
DNA Isolation
37.9K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
37.9K


