Open AccessSemiautomated Segmentation and Measurement of Cytoplasmic Vacuoles in a Neutrophil With General‐Purpose Image Analysis Software
Author(s) -
Mizukami Maki,
Yamada Misaki,
Fukui Sayaka,
Fujimoto Nao,
Yoshida Shigeru,
Kaga Sanae,
Obata Keiko,
Jin Shigeki,
Miwa Keiko,
Masauzi Nobuo
Publication year - 2016
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.21957
Subject(s) - cytoplasm , vacuole , segmentation , grey level , pixel , measure (data warehouse) , image (mathematics) , chemistry , physics , anatomy , microbiology and biotechnology , biology , optics , computer science , computer vision , biochemistry , data mining
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. Conclusion 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.