戊二醛和醋酸铀双重固定结合溅射/揭顶法能够实现细胞内脂肪酸的飞行时间二次离子质谱成像，并呈现与细胞器相对应的亚细胞分布。

Glutaraldehyde and uranyl acetate dual fixation combined sputtering/unroofing enables intracellular fatty acids TOF-SIMS imaging with organelle-corresponding subcellular distribution.

作者信息

Zhang Chi, Horikawa Makoto, Kahyo Tomoaki, Matsudaira Takaomi, Tanaka Tatsuya, Xu Lili, Takei Shiro, Setou Mitsutoshi

机构信息

Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.

International Mass Imaging Center, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.

出版信息

Microscopy (Oxf). 2022 Dec 8;71(6):324-333. doi: 10.1093/jmicro/dfac034.

DOI:10.1093/jmicro/dfac034

PMID:35762441

Abstract

Fatty acids (FAs) have diverse functions in cellular activities. The intracellular distribution of FAs is critical for their functions. Imaging of FAs by time-of-flight secondary ion mass spectrometry (TOF-SIMS) has been achieved. However, TOF-SIMS images of FAs so far do not have subcellular distribution due to inadequate sample preparation methods. In this study, we developed a chemical fixation method using glutaraldehyde (GA) with uranyl acetate (UA), which preserved cellular structure and intracellular FA distribution well. Combining GA+UA fixation with sputtering-based methods and unroofing-based methods, respectively, we successfully imaged intracellular lipids with the subcellular distribution.

摘要

脂肪酸（FAs）在细胞活动中具有多种功能。脂肪酸的细胞内分布对其功能至关重要。通过飞行时间二次离子质谱（TOF-SIMS）对脂肪酸进行成像已经实现。然而，由于样品制备方法不足，迄今为止脂肪酸的TOF-SIMS图像没有亚细胞分布。在本研究中，我们开发了一种使用戊二醛（GA）和醋酸铀（UA）的化学固定方法，该方法能很好地保留细胞结构和细胞内脂肪酸分布。分别将GA+UA固定与基于溅射的方法和基于去顶的方法相结合，我们成功地对具有亚细胞分布的细胞内脂质进行了成像。