title: Semi-Automatic Rating Method for Neutrophil Alkaline Phosphatase Activity.
abstract: BACKGROUND
  The neutrophil alkaline phosphatase (NAP) score is a valuable test for the diagnosis of myeloproliferative neoplasms, but it has still manually rated. Therefore, we developed a semi-automatic rating method using Photoshop®and Image-J, called NAP-PS-IJ.
  
  
  METHODS
  Neutrophil alkaline phosphatase staining was conducted with Tomonaga's method to films of peripheral blood taken from three healthy volunteers. At least 30 neutrophils with NAP scores from 0 to 5+ were observed and taken their images. From which the outer part of neutrophil was removed away with Image-J. These were binarized with two different procedures (P1 and P2) using Photoshop®. NAP-positive area (NAP-PA) and granule (NAP-PGC) were measured and counted with Image-J.
  
  
  RESULTS
  The NAP-PA in images binarized with P1 significantly (P < 0.05) differed between images with NAP scores from 0 to 3+ (group 1) and those from 4+ to 5+ (group 2). The original images in group 1 were binarized with P2. NAP-PGC of them significantly (P < 0.05) differed among all four NAP score groups. The mean NAP-PGC with NAP-PS-IJ indicated a good correlation (r = 0.92, P < 0.001) to results by human examiners.
  
  
  CONCLUSIONS
  The sensitivity and specificity of NAP-PS-IJ were 60% and 92%, which might be considered as a prototypic method for the full-automatic rating NAP score.
authors.affiliation: Department of Clinical Laboratory, NTT East Japan Sapporo Hospital, Sapporo, Japan.
authors.forename: Kanae
authors.initials: K
authors.lastname: Sugano
authors.affiliation: Department of Clinical Laboratory, Daiichi Kishimoto Medical Laboratory Center, Sapporo, Japan.
authors.forename: Kotomi
authors.initials: K
authors.lastname: Hashi
authors.affiliation: Department of Clinical Laboratory, Sapporo Hokuyu Hospital, Sapporo, Japan.
authors.forename: Misaki
authors.initials: M
authors.lastname: Goto
authors.affiliation: Department of Clinical Laboratory, Teine Keijinnkai Hospital, Sapporo, Japan.
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Nishi
authors.affiliation: Department of Clinical Laboratory, KKR Sapporo Medical Center, Sapporo, Japan.
authors.forename: Rie
authors.initials: R
authors.lastname: Maeda
authors.affiliation: Graduate school of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Keigo
authors.initials: K
authors.lastname: Kono
authors.affiliation: Graduate school of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Department of Medical laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Okada
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.|Department of Cardiovascular Regenerative Medicine, Osaka University Graduate School of Medicine, Suita, 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.
authors.forename: Taisei
authors.initials: T
authors.lastname: Mikami
authors.affiliation: Department of Medical laboratory Science, Faculty of Health Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Masauzi
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D001331:Automation; D057075:Enzyme Assays / Q000379:methods*; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009504:Neutrophils / Q000201:enzymology*; D055815:Young Adult
chemical_list: D000469:Alkaline Phosphatase
keywords: Image-J; Photoshop; Tomonaga's method; digital image processing; granule counting