Table 3.
Summary of quality control parameters for multiplex immunohistochemsitry analysis
| E-Cadherin Panel | Normal (N = 7) | IPF (N = 7) | Mann-Whitney U (Nonparametric) | Effect Size | ||
|---|---|---|---|---|---|---|
| Variable | Mean1 | SD | Mean2 | SD | p-value | |
| Number of ROIs in analysis | 116 | 31.5 | 92 | 37.9 | 0.259 | |
| Total number cell-sized objects | 193,341 | 43,830 | 156,428 | 59,257 | 0.209 | |
| Hoechst 33258 Positive Cells | 58,854 | 28,534 | 48,439 | 34,558 | 0.209 | |
| Avg Cell Area (μm2) | 126.4 | 8.0 | 125.1 | 4.8 | 0.535 | |
| Avg Cytoplasm Area (μm2) | 73.0 | 11.5 | 75.2 | 7.6 | 0.902 | |
| Avg Nucleus Area (μm2) | 53.4 | 5.0 | 49.9 | 5.1 | 0.259 | |
| Avg Nucleus Perimeter (μm) | 34.4 | 2.1 | 33.7 | 1.8 | 0.456 | |
| Avg Nucleus Roundness | 0.7 | 0.005 | 0.7 | 0.007 | 0.001** | |
| Area Analyzed (μm2) | 5E+ 07 | 2E+ 07 | 4E+ 07 | 2E+ 07 | 0.383 | −2.073 |
| CD45 Panel | ||||||
| Normal (N = 7) | IPF (N = 7) | Mann-Whitney U (Nonparametric) | Effect Size | |||
| Variable | Mean1 | SD | Mean2 | SD | p-value | |
| Number of ROIs in analysis | 111 | 32.4 | 91 | 33.5 | 0.318 | |
| Total number cell-sized objects | 186,596 | 52,479 | 153,307 | 53,860 | 0.318 | |
| Hoechst 33258 Positive Cells | 52,770 | 31,528 | 41,252 | 31,069 | 0.456 | |
| Avg Cell Area (μm2) | 125.0 | 6.9 | 123.6 | 5.9 | 0.902 | |
| Avg Cytoplasm Area (μm2) | 71.9 | 10.4 | 73.0 | 8.8 | 1.000 | |
| Avg Nucleus Area (μm2) | 53.0 | 4.8 | 50.6 | 4.3 | 0.383 | |
| Avg Nucleus Perimeter (μm) | 34.6 | 2.1 | 34.1 | 1.8 | 0.805 | |
| Avg Nucleus Roundness | 0.7 | 0.005 | 0.7 | 0.006 | 0.001** | −2.309 |
| Area Analyzed (μm2) | 5E+ 07 | 2E+ 07 | 5E+ 07 | 2E+ 07 | 0.646 | |
Quality control parameters were quantified from the regions of interest (ROIs) used in the multiplex analysis presented in Table 3 and Figs. 4 and 5. The parameters of normal (n = 7) and idiopathic pulmonary fibrosis (IPF, n = 7) lung tissue was compared using Mann-Whitney U test. Effect size (Cohen’s D) was calculated for comparisons that were significantly different (*p < 0.05, **p < 0.01). Avg, average