Porter Corey M, Truttmann Matthias C
bioRxiv. 2023 Sep 22:2023.09.19.558493. doi: 10.1101/2023.09.19.558493.
Comprehensive characterization of protein networks in mounted brain tissue represents a major challenge in brain and neurodegenerative disease research. In this study, we develop a simple staining method, called TSWIFT, to iteratively stain pre-mounted formalin fixed, paraffin embedded (FFPE) brain sections, thus enabling high-dimensional sample phenotyping. We show that TSWIFT conserves tissue architecture and allows for relabeling a single mounted FFPE sample more than 10 times, even after prolonged storage at 4 °C. Using TSWIFT, we profile the abundance and localization of the HSP70 family chaperones HSC70 (HSPA8) and BiP (HSPA5) in mounted human brain tissue. Our results establish TSWIFT as an efficient method to obtain integrated high-dimensional knowledge of cellular proteomes by analyzing mounted FFPE human brain tissue.
对已封片的脑组织中的蛋白质网络进行全面表征是脑和神经退行性疾病研究中的一项重大挑战。在本研究中，我们开发了一种名为TSWIFT的简单染色方法，用于对预先封片的福尔马林固定、石蜡包埋（FFPE）脑切片进行反复染色，从而实现高维样本表型分析。我们表明，TSWIFT可保留组织结构，即使在4°C长期保存后，也能对单个已封片的FFPE样本进行超过10次的重新标记。使用TSWIFT，我们对已封片的人类脑组织中HSP70家族伴侣蛋白HSC70（HSPA8）和BiP（HSPA5）的丰度和定位进行了分析。我们的结果表明，TSWIFT是一种通过分析已封片的FFPE人类脑组织来获得细胞蛋白质组综合高维知识的有效方法。
