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Table 3 Summary of quality control parameters for multiplex immunohistochemsitry analysis

From: CXCR4+ cells are increased in lung tissue of patients with idiopathic pulmonary fibrosis

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

 
  1. 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