Glutaraldehyde and uranyl acetate dual fixation combined sputtering/unroofing enables intracellular fatty acids TOF-SIMS imaging with organelle-corresponding subcellular distribution. (28th June 2022)

• Record Type: Journal Article
• Authors: Zhang, Chi; Horikawa, Makoto; Kahyo, Tomoaki; Matsudaira, Takaomi; Tanaka, Tatsuya; Xu, Lili; Takei, Shiro; Setou, Mitsutoshi
• Is Part Of: Microscopy. Volume 71:Number 6(2022)
• Page Start: 324
• Page End: 333
• Publication Date: 2022-06-28
• Subjects: TOF-SIMS -- single-cell imaging -- intracellular FA imaging -- chemical fixation -- TOF-SIMS sample preparation -- FA subcellular imaging; Microscopy -- Periodicals; 502.825
• DOI: 10.1093/jmicro/dfac034
• Languages: English
• ISSNs: 2050-5698

Abstract

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.