title: Three-Dimensional Image of Cleavage Bodies in Nuclei Is Configured Using Gas Cluster Ion Beam with Time-of-Flight Secondary Ion Mass Spectrometry.
abstract: Structural variations of DNA in nuclei are deeply related with development, aging, and diseases through transcriptional regulation. In order to bare cross sections of samples maintaining sub-micron structures, an Ar2500(+)-gas cluster ion beam (GCIB) sputter was recently engineered. By introducing GCIB sputter to time-of-flight secondary ion mass spectrometry (TOF-SIMS), we analyzed the 3D configuration and chemical composition of subnuclear structures of pyramidal cells in the CA2 region in mouse brain hippocampus. Depth profiles of chemicals were analyzed as 3D distributions by combining topographic analyses. Signals corresponding to anions such as CN(-) and PO3(-) were distributed characteristically in the shape of cell organelles. CN(-) signals overlapped DAPI fluorescence signals corresponding to nuclei. The clusters shown by PO3(-) and those of adenine ions were colocalized inside nuclei revealed by the 3D reconstruction. Taking into account their size and their number in each nucleus, those clusters could be in the cleavage bodies, which are a kind of intranuclear structure.
authors.affiliation: Dept of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Noritaka
authors.initials: N
authors.lastname: Masaki
authors.affiliation: ULVAC-PHI, 370 Enzo, Chigasaki, Kanagawa 253-8522, Japan.
authors.forename: Itsuko
authors.initials: I
authors.lastname: Ishizaki
authors.affiliation: Dept of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hayasaka
authors.affiliation: Physical Electronics, 18725 Lake Drive East, Chanhassen, MN 55317, USA.
authors.forename: Gregory L
authors.initials: GL
authors.lastname: Fisher
authors.affiliation: ULVAC-PHI, 370 Enzo, Chigasaki, Kanagawa 253-8522, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Sanada
authors.affiliation: Image Processing Research Team, Center for Advanced Photonics, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Yokota
authors.affiliation: Dept of Cell Biology and Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Setou
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice