Zhang Chi, Horikawa Makoto, Kahyo Tomoaki, Matsudaira Takaomi, Tanaka Tatsuya, Xu Lili, Takei Shiro, Setou Mitsutoshi
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.
Microscopy (Oxf). 2022 Dec 8;71(6):324-333. doi: 10.1093/jmicro/dfac034.
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.
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。
