title: Semiautomated Segmentation and Measurement of Cytoplasmic Vacuoles in a Neutrophil With General-Purpose Image Analysis Software.
abstract: BACKGROUND
  Morphological observation of blood or marrow film is still described nonquantitatively. We developed a semiautomatic method for segmenting vacuoles from the cytoplasm using Photoshop (PS) and Image-J (IJ), called PS-IJ, and measured the relative entire cell area (rECA) and relative areas of vacuoles (rAV) in the cytoplasm of neutrophil with PS-IJ.
  
  
  METHODS
  Whole-blood samples were stored at 4°C with ethylenediaminetetraacetate and in two different preserving manners (P1 and P2). Color-tone intensity levels of neutrophil images were semiautomatically compensated using PS, and then vacuole portions were automatically segmented by IJ. The rAV and rECA were measured by counting pixels by IJ. For evaluating the accuracy in segmentations of vacuoles with PS-IJ, the rAV/rECA ratios calculated with results from PS-IJ were compared with those calculated with human eye and IJ (HE-IJ).
  
  
  RESULTS
  The rECA and rAV/ in P1 significantly (P < 0.05, P < 0.05) were enlarged and increased, but did not significantly (P = 0.46, P = 0.21) change in P2. The rAV/rECA ratios by PS-IJ were significantly correlated (r = 0.90, P < 0.01) with those by HE-IJ.
  
  
  CONCLUSIONS
  PS-IJ method can successfully segment vacuoles and measure the rAV and rECA, becoming a useful tool for quantitative description of morphological observation of blood and marrow film.
authors.affiliation: Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan.
authors.forename: Maki
authors.initials: M
authors.lastname: Mizukami
authors.affiliation: Department of Clinical Laboratory, Japanese Red Cross Medical Center, Tokyo, Japan.
authors.forename: Misaki
authors.initials: M
authors.lastname: Yamada
authors.affiliation: Department of Clinical Laboratory, Hokkaido Industrial Health Management Fund, Sapporo, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Fukui
authors.affiliation: Department of Clinical Laboratory, Hokkaido Industrial Health Management Fund, Sapporo, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Fujimoto
authors.affiliation: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Yoshida
authors.affiliation: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Sanae
authors.initials: S
authors.lastname: Kaga
authors.affiliation: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Obata
authors.affiliation: Department of Fundamental Nursing, Facalty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Jin
authors.affiliation: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.|Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Osaka, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Miwa
authors.affiliation: Department of Medical Laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan. nmasauzi@hs.hokudai.ac.jp.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Masauzi
mesh_terms: D000704:Analysis of Variance; D001769:Blood; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008297:Male; D009504:Neutrophils / Q000166:cytology*; D012984:Software; D013997:Time Factors; D014617:Vacuoles / Q000473:pathology / Q000648:ultrastructure*
keywords: Image-J; Photoshop; area; differentiation; quantitative