以莱克为媒介,时间高效,高产分类不同的形态完整的脏片段
Jessica Roskosch1, Uyen Huynh-Do1, Stefan Rudloff2
1Division of Nephrology and Hypertension, University of Bern and University Hospital Bern, Freiburgstrasse 15, CH-3010, Bern, Switzerland.
Pflugers Archiv : European journal of physiology
|December 13, 2023
概括
研究人员开发了一种新的方法,使用光学蛋白和流量分类来快速分离纯脏脏片段. 这种技术保留了形态,并产生了大量的蛋白质和RNA,用于高级脏研究.
科学领域:
- 腎臟病學 (nephrology) 是一種醫學專業.
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 脏的脏是复杂的结构,有不同的部分,对于过和再吸收至关重要.
- 传统的用于隔离脏片段的方法是繁的,耗时的,并且产生有限的材料.
- 需要高效和高产量的技术来获得用于研究的纯脏片段.
研究的目的:
- 开发一种新,快速,经济高效的方法,同时分离多个脏脏片段.
- 为下游分析保留孤立脏片段的3D形态.
- 为了从小鼠和人类脏样本中高纯度地分离脏片段.
主要方法:
- 组合低成本的化物-合性莱克或聚合素 (Flaggs) 与流量分类.
- 使用特定标志 (SNA-Cy3,LTL-FITC,SBA-PB) 来对脏片段进行差异标记.
- 采用200μm喷嘴和5psi进行分类,分类质体,近端卷状管体和远端卷状管体.
- 通过双重标记识别的连接管道/收集管道和缺乏标记的厚厚的上升四肢.
主要成果:
- 在不到3小时的时间内,同时分离不同的脏片段,保持3D形态.
- 每个分类段实现了蛋白质 (37-521 ng/s) 和RNA (0.71-16.71 ng/s) 的高产量.
- 已证明分类段的高纯度,mRNA表达水平的中位丰富度为96.1倍.
结论:
- 新的旗和流量分类方法是简单的,具有成本效益,并且广泛适用.
- 这种技术提供了高产量的纯,形态完整的管材料.
- 该方法有可能显著推进脏细分特定研究.
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