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用过器辅助扩张蛋白质学用于FFPE组织样本中单个细胞和有机体的空间分析

Zhen Dong^1,2,3, Chunlong Wu^1,2,3, Jiayi Chen^1,2,3

¹Affiliated Hangzhou First People's Hospital, State Key Laboratory of Medical Proteomics, School of Medicine, Westlake University, Hangzhou, China.

Nature protocols

|October 9, 2025

概括

过器辅助扩展蛋白质组学 (FAXP) 能够在FFPE组织中实现高分辨率的空间蛋白质组学. 这种方法保留了空间特征和蛋白质完整性,推进了癌症和神经退行性疾病研究.

科学领域:

蛋白质组学是指蛋白质组学.
生物技术是生物技术.
历史学历史学历史学

背景情况:

固定在形式素中,嵌入式抛 (FFPE) 组织对于研究至关重要,但对于高分辨率空间蛋白质组学具有挑战性.
现有的方法通常在分析过程中难以保留空间信息和蛋白质完整性.

研究的目的:

引入过器辅助扩张蛋白质组学 (FAXP),这是FFPE组织的新型空间蛋白质组学方法.
为了使同位素组织扩张和强大的蛋白质保留,同时保持空间保真.

主要方法:

FAXP将基于水凝的组织扩张与FFPE样本的质谱相结合.
工作流包括脱,蛋白质定,水凝嵌入,同位素膨胀和过器辅助的凝消化.
激光捕捉微解剖用于单细胞和亚细胞空间蛋白质组学.

主要成果:

即使在富含ECM的样本中,FFPE组织的线性扩张达到5倍,扭曲率低于6%,即使在富含ECM的样本中.
从单个小鼠肝核中平均发现了2,368种蛋白质,从单个细胞中发现了3,312种蛋白质.
已证明与各种组织类型的兼容性以及与免疫染工作流程的无集成.

结论:

FAXP提供了FFPE组织的全面,可复制和敏感的蛋白质基因特征.
~27小时的工作流可供具有中级专业知识的研究人员访问.
FAXP准备推进癌症异质性,神经退行性疾病和细胞微环境的研究.

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