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Additionally manual quantification also generates bias by different observers, as shown in this study.
However, manual quantification of PMN is operator-dependent and can delay the diagnosis and in fine antibiotic administration.
Manual quantification of 5 cells per embryo of each genotype (wild type, embryos expressing Rab5DN, Rab11DN or both simultaneously in the AS) was performed using ImageJ.
We show here in box-plots that the number of apoptotic neurons increases between stages 13 and 17 in Drosophila embryos (Figure 6), consistently with a previous report of manual quantification of Caspase stained cells [11].
Spatial 3D reconstructions integrate spatial coordinates from two cameras (positioned as in Fig. 1) within an X,Y,Z-scatter plot (Fig. 5), again showing how video-tracking software detects complex anxiety-like behaviors previously limited to manual quantification (see Fig. 6 for other generated 3D traces, and Supplementary Material for details).
These skin biopsies were analyzed by manual quantification of immunohistochemical double-staining and quantitative RT-PCR.
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In addition, the comparison of the derived information with the on-site manual quantifications revealed the potentiality of the QUAV and multi-stage system for future practice.
To complement the manual quantifications of a restricted number of AS cells over time, we also performed a semi-automated analysis of the entire AS tissue in embryos at the beginning of DC.
For joint effusion volume, Pearson's correlation was used for the comparison between automated and manual quantifications, and between automated quantification and arthrocentesis.
Manual quantifications were also performed for co-localization event between Jen1-GFP with markers of internal compartments (Sec7-mCh and Vps17-mCh).
Although the developed automated and manual quantifications used different segmentation processes and a slightly different volume calculation (subvoxel versus voxel size), comparison revealed high statistical correlation, suggesting that the developed automated system is reliable and provides precise joint effusion volume.
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