Identification of New Rat Bone Marrow-Derived Population of Very Small Stem Cell with Oct-4A and Nanog Expression by Flow Cytometric Platforms

Figure 4

Quantitative analysis of morphological features of selected rat BM populations by ImageStream X system. (a) Quantitative analysis of cell size, nuclear to cytoplasmic (N/C) ratio, and cytoplasmic area of rVSELs when compared to other cell types. Analysis was performed based on the collected images of following rat BM populations: (i) rVSELs: CD45⁻/Lin⁻/CD106⁺ cells expressing Oct-4A or Nanog (upper and lower table, resp.); (ii) rHSCs: CD45⁺/Lin⁻/CD106⁺ cells with no expression of Oct-4A and Nanog, and (iii) TNCs (gated as singlets during the analysis). Analyses were performed with IDEAS software (Amnis Corp.). Data are shown as average values of each indicated feature within the cell population (mean ± SD). (b) Analytical approach for masking and feature calculation by IDEAS. Images show brightfield (BF), nuclear (Hoe), and combined (BF-Hoe) images of one example rat BM cell as well as exact masks (cyan) and features (formulas based on the defined masks) that were optimized and used for the analysis shown in (a). Cell size was expressed as minor cell axis computed within the mask covering BF image, while cytoplasmic area and N/C ratio were calculated based on the areas of BF and nuclear images (as indicated in red boxes). All masks and features including their nomenclature represent standard parameters provided by IDEAS software (Amnis Corp.)

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