Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Representative dot plots on the C6 Plus flow cytometer showing comparison of data acquisition utilizing unadjusted and adjusted settings with data acquisition images of positive sample with ( a ) unadjusted flame shaped gate settings on C6 Plus flow cytometer (FSC-A vs SSC-A), ( b ) unadjusted settings on C6 Plus flow cytometer (FSC-A vs PI), ( c ) unadjusted settings on C6 Plus flow cytometer (FITC-A vs PI), ( d ) adjusted settings on C6 Plus flow cytometer (FSC-A vs SSC-A) including the sperm and the apoptotic bodies. ( e ) Data plots of positive sperm sample with the adjusted settings on C6 Plus flow cytometer (FSC-A vs PI) including the PI-positive events within the gate and excluding the M450 apoptotic bodies. ( f ) Data acquisition images of positive sperm sample with the adjusted settings on C6 Plus flow cytometer (FITC-A vs PI): the TUNEL/PI-positive events are depicted in the right upper quadrant and this estimates the TUNEL positivity. FITC-A: fluorescein isothiocynate-area; SSC-A: side scatter area; PI: propidium iodide; UL: upper left; UR: upper right; LL: lower left; LR: lower right; P1: gate P1; P3: gate P3; Q2: quadrant 2.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques: Flow Cytometry, TUNEL Assay

Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Dot plots obtained on the C6 Plus with the modified or adjusted gates for negative control sperm sample. The first plot a depicts the FSC-A versus SSC-A plot with flame shaped gate that captures the spermatozoa and apoptotic bodies, plot b is a PI-A/FSC-A plot with PI positive events. This gate includes all spermatozoa and excludes the M450 apoptotic bodies, ( c ) is the PI-A/FITC-A plot for the negative sperm control which is used to set the quadrants, ( d ) depicts the PI-A versus FITC-A gate plot of a negative test sample. The positive sperm sample was run in duplicate, and shows SDF of ( e ) 36.7% and ( f ) 36.1 %. The SDF values of the test sample run in duplicate reflect the very low intra-assay variability. FITC-A: fluorescein isothiocynate-area; SSC-A: side scatter-area; PI: propidium iodide; SDF: sperm DNA fragmentation; UL: upper left; UR: upper right; LL: lower left; LR: lower right; P2: gate P2; P4: gate P4; Q1: quadrant 1.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques: Modification, Negative Control, Intra Assay

Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Percentage chart showing extent of DNA fragmentation in ( a ) measurements without DNA fragmentation (orange) and DNA fragmentation (blue) on C6 Plus without adjustment when compared with C6 standard. In measurements without DNA fragmentation, it could not differentiate the false-positive measurements and all were identified as true-negative measurements. However, in measurements with DNA fragmentation, it incorrectly identified 40% of measurements as false negative and only 60% as true positive, ( b ) same measurements on C6 Plus after adjustment. The number of false -positive measurements without DNA fragmentation was significantly low. Majority of the measurements were correctly identified as true positive in case of DNA fragmentation. TN: true negative; FP: false positive; FN: false negative; TP: true positive.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques:

Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Receiver operating characteristic curve showing sensitivity, specificity, and cutoff for C6 Plus ( a ) in unadjusted SDF and ( b ) after adjustment with the standard C6 flow cytometer. AUC: area under curve; SDF: sperm DNA fragmentation.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques: Flow Cytometry

Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Bland-Altman plots for ( a ) unadjusted C6 Plus versus C6 standardized and ( b ) adjusted C6 Plus versus C6 standardized. SDF: sperm DNA fragmentation; s.d.: standard deviation.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques: Standard Deviation

Journal: Asian Journal of Andrology

Article Title: Critical evaluation of two models of flow cytometers for the assessment of sperm DNA fragmentation: an appeal for performance verification

doi: 10.4103/aja.aja_109_18

Figure Lengend Snippet: Passing-Bablok regression analysis showing ( a ) unadjusted C6 Plus versus C6 standardized (the wider deviation of the values from one another can clearly be seen), and ( b ) adjusted C6 Plus versus C6 standardized (the deviation in the values is much narrower). Red dashed line shows the confidence interval of the regression line expected averages of SDF; red dotted line is the identity line and blue is the regression line of the measured averages. SDF: sperm DNA fragmentation; s.d.: standard deviation.

Article Snippet: Comparison of the C6 and C6 Plus flow cytometer was done with the MedCalc Statistical Software, version 18.2.1 (MedCalc Software bvba, Ostend, Belgium; https://www.medcalc.org; 2017 ).

Techniques: Standard Deviation