使用超离心法从固体介质培养物中分离真菌细胞外囊泡的协议
Alicia C Piffer1, Flavia C G Reis2, Frédérique Moyrand1
1Institut Pasteur, Université Paris Cité, Unité Biologie des ARN des Pathogènes Fongiques, Département de Mycologie, 75015 Paris, France.
STAR protocols
|May 21, 2024
概括
本协议详细介绍了分离和分析真菌细胞外囊泡 (EV) 的方法,这些方法对于了解真菌病原体至关重要. 这些技术使得高通量选和集中样品准备能够用于各种下游应用.
科学领域:
- 菌类学 菌类学是指菌类学.
- 细胞生物学 细胞生物学
- 生物技术是生物技术.
背景情况:
- 细胞外囊泡 (EVs) 在真菌生物学中越来越多地被识别,包括在人类致病物种中.
- 了解真菌EV对于开发新型诊断和治疗策略至关重要.
研究的目的:
- 为分离和分析真菌细胞外囊泡 (EVs) 提出两个不同的,优化的协议.
- 为研究人员提供一个全面的指南与真菌EVs工作,促进进一步调查他们的角色.
主要方法:
- 描述一种高通量选方法,用于快速隔离真菌EV.
- 获取密度梯度超离心适合密度梯度超离心的缩真菌EV样本的详细协议.
- 关于分析试验的指导,包括纳米流细胞计和纳米粒子跟踪分析,用于EV的表征.
主要成果:
- 提出的方法允许有效地从各种真菌物种中分离真菌EVs.
- 分析技术可以准确测量EV尺寸分布和度.
- 电动汽车悬架适用于广泛的下游应用,包括显微镜和组成分析.
结论:
- 开发的协议为真菌EV研究提供了强大而多样化的方法.
- 这些方法将有助于推进对真菌EVs,它们的生物发生及其在健康和疾病中的功能作用的研究.
- 标准化隔离和分析技术对于不同真菌病原体的比较研究至关重要.
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