提高细胞保留,尤其在细胞数量较少的情况下
减少细胞压力,提高细胞活力
减少碎片和细胞团聚
提高细胞群的分辨率
省时省力,通量很高
即将举行的网络研讨会:实现无需离心的流式细胞术样品制备工作流程的标准化
层流洗涤技术适用于相信定量流式细胞术对细胞治疗、免疫肿瘤学和其他细胞分析很重要的领域至关重要的科学家。
单细胞基因组学和蛋白质基因组学正在迅速发展。Nature Methods将多模态组学评为2019年年度方法。随着越来越多的科学家使用单细胞技术,他们发现样品制备对于产生高质量数据至关重要。
细胞团聚、细胞保留率低和残留核酸是影响单细胞测序和蛋白质基因组学的几个重要原因,与此同时,离心还会对样品产生机械压力,从而导致结果出现偏差。
样品制备在单细胞基因组学和蛋白质基因组学中极其重要,尤其是对于单细胞基因表达和免疫分析。当mRNA转录物被编码时,mRNA从破碎的细胞中溢出(增加背景)或mRNA降解(减少信号)会显著影响测量结果。与流式细胞术不同,在单细胞测序中不可能排除碎片或细胞群。细胞制备必须尽可能干净且为单细胞。
10X Genomics的样品制备证明了用于单细胞RNA测序的新鲜冷冻人外周血单核细胞使用离心来清洗细胞需要注意“不要过度离心”,因为离心速度会破坏细胞的完整性和细胞活力。一项发表在Nature Methods上的研究表明,单细胞分离程序会引起细胞内转录组范围的变化。因此,如果有一种更温和的方法来清洗细胞,将会吸引大批科学家来使用新技术。
使用层流洗涤系统可以通过更少的手动操作来节省时间。在没有非自然离心力的情况下,温和地洗涤细胞可以保持细胞的完整性和细胞活力,同时确保基因或蛋白质表达的变化是生物学上的变化,而不是工作流程产生的偏差。
层流洗涤技术已被加入官方TotalSeq™方案。它简化了样品处理流程,同时实现了高通量。
“Curiox (Laminar Wash) really helps retain high numbers of cells”
Christoph Eberle 博士,查尔斯河实验室Antonino Montalbano博士,ImmunAI
用于制备用于流式细胞术分析的单细胞悬液的免疫染色工作流程的比较。层流洗涤技术在洗涤方面同样有效,且无需使用离心机。
提高细胞保留,尤其在细胞数量较少的情况下
减少细胞压力,提高细胞活力
减少碎片和细胞团聚
提高细胞群的分辨率
省时省力,通量很高
Single-Cell Transcriptomic Analyses Define Distinct Peripheral B Cell Subsets and Discrete Development Pathways.
Stewart, et al. (2021) Front. Immunol.
图 1:层流洗涤产生更好的细胞散列结果
使用 Curiox 层流清洗系统处理的样品产生更少的双峰、更少的阴性结果和更多的单峰。数据由 ImmunAI 提供,改编自Antonio Montalbano 博士与 Curiox Biosystems 的网络研讨会。
图2:层流洗涤保留更多细胞
与传统的基于离心机的方法相比,层流洗涤始终保留更细胞。对于细胞数量非常少的样品尤其如此。数据由 ImmuzunAI 提供,改编自Antonio Montalbano 博士与 Curiox Biosystems 的网络研讨会。
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