scholarly journals Comparing flow cytometry and fluorescence microscopy for analyzing human sperm DNA fragmentation by TUNEL labeling

2008 ◽  
Vol 73A (9) ◽  
pp. 785-787 ◽  
Author(s):  
Monica Muratori ◽  
Gianni Forti ◽  
Elisabetta Baldi
Keyword(s):  
Flow Cytometry ◽  
Fluorescence Microscopy ◽  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

scholarly journals Human sperm DNA fragmentation: Correlation of TUNEL results as assessed by flow cytometry and optical microscopy

2007 ◽  
Vol 71A (12) ◽  
pp. 1011-1018 ◽  
Author(s):  
David Domínguez-Fandos ◽  
María Isabel Camejo ◽  
José Luis Ballescà ◽  
Rafael Oliva
Keyword(s):  
Flow Cytometry ◽  
Optical Microscopy ◽  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

Use of laptop computers connected to internet through Wi-Fi decreases human sperm motility and increases sperm DNA fragmentation

2012 ◽  
Vol 97 (1) ◽  
pp. 39-45.e2 ◽  
Author(s):  
Conrado Avendaño ◽  
Ariela Mata ◽  
César A. Sanchez Sarmiento ◽  
Gustavo F. Doncel
Keyword(s):  
Dna Fragmentation ◽  
Sperm Motility ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Laptop Computers ◽  
Sperm Dna

DETERMINATION OF THE HUMAN SPERM DNA FRAGMENTATION INDEX

Vestnik ◽  
2021 ◽  
pp. 226-232
Author(s):  
К. К. Тлеуханов ◽  
Н. А. Алтыбаева ◽  
М. К. Отарбаев ◽  
Е. М. Тойшибеков ◽  
А. А. Тлеуханова
Keyword(s):  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

В статье представлены собранные данные о методах устранения повышенной частоты фрагментации ДНК у сперматозоидов, которые в некоторых исследованиях подтверждают, что введение антиоксидантов может снизить уровень фрагментации ДНК у сперматозоидов. The article presents collected data on methods of eliminating the increased frequency of DNA fragmentation in spermatozoa, which in some studies confirm that the introduction of antioxidants can reduce the level of DNA fragmentation in spermatozoa.

scholarly journals Critical Aspects of Detection of Sperm DNA Fragmentation by Tunel/Flow Cytometry

2010 ◽  
Vol 56 (4) ◽  
pp. 277-285 ◽  
Author(s):  
Monica Muratori ◽  
Lara Tamburrino ◽  
Sara Marchiani ◽  
Carmen Guido ◽  
Gianni Forti ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Fragmentation ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Critical Aspects

scholarly journals 265 SPERM DNA FRAGMENTATION AND PREGNANCY OUTCOME

2005 ◽  
Vol 17 (2) ◽  
pp. 282
Author(s):  
D. Evenson
Keyword(s):  
Flow Cytometry ◽  
Pregnancy Outcome ◽  
Dna Fragmentation ◽  
Dna Strand Breaks ◽  
Sperm Dna Fragmentation ◽  
Strand Breaks ◽  
Threshold Levels ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Dna Strand

Sperm DNA integrity is obviously important for normal embryo development and pregnancy outcome. Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. The Sperm Chromatin Structure Assay (SCSA) treats sperm with low pH to denature DNA at the sites of DNA strand breaks, followed by acridine orange (AO) staining of green for native DNA and red for denatured DNA, as measured by flow cytometry (FCM), as well as % sperm with high DNA stainability (HDS: immature sperm with intact DNA related to decreased fertilization rates). FCM-sorted sperm from each SCSA-defined population (normal, moderate, and high DNA fragmentation and HDS sperm) show that the moderate DNA fragmentation index (DFI) population has the same image analysis characteristics as normal sperm without significant comets. Thus, an ICSI technician is not likely to differentiate between a normal and a moderate DFI sperm. The TUNEL assay uses an enzyme to add a fluorochrome-labeled base to a 3′-OH broken DNA strand. Both light microscopy and flow cytometry are used for measuring the % and extent of DNA fragmentation but cannot measure the level of HDS. For the COMET assay, sperm are suspended in an electrophoretic gel, placed on a glass microscope slide, digested with proteases and RNAse, subjected to an electric field, and then stained with a DNA dye. The % of comet positive sperm is scored, but the extent of fragmentation is difficult to define and the % HDS cannot be determined. Small pieces of fragmented DNA migrate in the gel forming a “comet.” All three methods have been used for both research and clinical diagnosis and as prognosis for livestock (bulls, boars, rams, stallions) and humans. Light microscope techniques suffer from a lack of statistical soundness needed for clinical decisions as well as present a potential bias in selection of sperm for measurements. Due to the thousands of sperm randomly selected for flow cytometry measurements, the data are statistically robust. Data from all three kinds of measurements in over a hundred manuscripts clearly show that sperm DNA fragmentation has a negative impact on embryo growth and pregnancy. Infertile animals may have nearly all of the sperm with fragmented DNA. Fertility ratings in bulls and boars are clearly related to the percent and extent of DNA fragmentation. Threshold levels for fertile/sub fertile/infertile differ for different species. Likewise different methods/laboratories have suggested various threshold levels to characterize a man with a highly fertile to low/very poor potential. The range of sperm with fragmented DNA is from ∼2% to 100%. The SCSA method has defined a 27–30% DFI as the point in which a man is placed into a statistical category of taking a longer time to achieve in vivo pregnancy, more intrauterine insemination and routine IVF cycles, or no pregnancy. Current data suggest that ICSI may help overcome the diminished pregnancy prognosis with high DFI over the other ART or natural methods.

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