Stain-free interferometric phase microscopy correlation with DNA fragmentation stain in human spermatozoa

2018 ◽  
Vol 11 (11) ◽  
pp. e201800137 ◽  
Author(s):  
Itay Barnea ◽  
Lidor Karako ◽  
Simcha K. Mirsky ◽  
Mattan Levi ◽  
Michal Balberg ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Human Spermatozoa ◽  
Phase Microscopy ◽  
Interferometric Phase

scholarly journals Potential effect of tobacco cigarettes smoking on global DNA methylation status and protamines transcripts in human spermatozoa

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Mohammed M. Laqqan ◽  
Maged M. Yassin
Keyword(s):  
Dna Methylation ◽  
Cigarette Smoking ◽  
Dna Fragmentation ◽  
Methylation Status ◽  
Human Spermatozoa ◽  
Global Dna Methylation ◽  
Transcription Level ◽  
Heavy Smokers ◽  
Protamine 1 ◽  
Protamine 2

Abstract Background Epigenetics refers to an alteration in gene expression without alteration in the sequence of DNA and this process may be affected by environmental factors and lifestyle like cigarette smoking. This study was designed to evaluate the potential effect of cigarette smoking on the global DNA methylation status and the transcription level of protamine 1 and protamine 2 in human spermatozoa. A total of 188 semen samples were collected from men with a mean age of 34.9 ± 5.8 years old (98 heavy smokers and 90 non-smokers). The DNA and RNA were isolated from purified spermatozoa, then the status of global DNA methylation and the transcription level of protamine 1 and protamine 2 were evaluated using ELISA and qPCR, respectively. The chromatin non-condensation and DNA fragmentation in human spermatozoa were evaluated using chromomycin A3 staining and TUNEL assay, respectively. Results A significant increase has been found in the status of global DNA methylation in spermatozoa of heavy smokers compared to non-smokers (7.69 ± 0.69 ng/μl vs. 4.90 ± 0.40 ng/μl, P < 0.001). Additionally, a significant reduction has been found in transcription level of protamine 1 (25.49 ± 0.31 vs. 23.94 ± 0.40, P < 0.001) and protamine 2 (28.27 ± 0.39 vs. 23.45 ± 0.30, P < 0.001) in heavy smokers. A downregulation has been found in the transcription level of protamine 1 and protamine 2 with a fold change of 0.497 and 0.047, respectively. A significant increase has been shown in the level of DNA fragmentation and chromatin non-condensation in heavy smokers compared to non-smokers (P < 0.001). On the other hand, a significant positive correlation has been found between sperm chromatin non-condensation, sperm DNA fragmentation, transcription level of protamine 1, transcription level of protamine 2, and global DNA methylation status (r = 0.304, P < 0.001; r = 0.399, P < 0.001; r = 0.216, P = 0.003; r = 0.494, P < 0.001, respectively). Conclusion Tobacco cigarette smoking has a potential influence on the global DNA methylation and the transcription level of protamine genes in human spermatozoa, and consequently, affect negatively on the semen parameters.

scholarly journals Whole-cell-analysis of live cardiomyocytes using wide-field interferometric phase microscopy

2010 ◽  
Vol 1 (2) ◽  
pp. 706 ◽  
Author(s):  
Natan T. Shaked ◽  
Lisa L. Satterwhite ◽  
Nenad Bursac ◽  
Adam Wax
Keyword(s):  
Wide Field ◽  
Cell Analysis ◽  
Whole Cell ◽  
Phase Microscopy ◽  
Interferometric Phase

scholarly journals DNA fragmentation of human spermatozoa: Simple assessment of single‐ and double‐strand DNA breaks and their respective dynamic behavioral response

Andrology ◽  
2020 ◽  
Vol 8 (5) ◽  
pp. 1287-1303
Author(s):  
Ana Tímermans ◽  
Rosana Vázquez ◽  
Fátima Otero ◽  
Jaime Gosálvez ◽  
Stephen Johnston ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Behavioral Response ◽  
Human Spermatozoa ◽  
Dna Breaks ◽  
Double Strand Dna Breaks ◽  
Double Strand Dna

scholarly journals Differences in nuclear DNA fragmentation and mitochondrial integrity of semen and prepared human spermatozoa

2000 ◽  
Vol 15 (7) ◽  
pp. 1552-1561 ◽  
Author(s):  
Eilish T. Donnelly ◽  
Michael O'Connell ◽  
Neil McClure ◽  
Sheena E.M. Lewis
Keyword(s):  
Dna Fragmentation ◽  
Nuclear Dna ◽  
Human Spermatozoa ◽  
Mitochondrial Integrity ◽  
Nuclear Dna Fragmentation

ITIA: tripling the field-of-view in off-axis interferometric phase microscopy

2014 ◽  
Author(s):  
Irena Frenklach ◽  
Pinhas Girshovitz ◽  
Natan T. Shaked
Keyword(s):  
Field Of View ◽  
Phase Microscopy ◽  
Interferometric Phase
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