PMID- 35532241
OWN - NLM
STAT- MEDLINE
DCOM- 20221229
LR  - 20221229
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 182
DP  - 2022 Apr 21
TI  - Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional 
      Defects.
LID - 10.3791/63790 [doi]
AB  - The conventional semen parameter analysis is widely used to assess male 
      fertility. However, studies have found that ~15% of infertile patients show no 
      abnormalities in conventional semen parameters. Additional technologies are 
      needed to explain the idiopathic infertility and detect subtle sperm defects. 
      Currently, biomarkers of sperm function, including sperm apoptosis, mitochondrial 
      membrane potential (MMP), and DNA damage, reveal sperm physiology at the 
      molecular level and are capable of predicting male fertility. With flow cytometry 
      (FCM) techniques, each of these markers can be rapidly, accurately, and precisely 
      measured in human semen samples, but time costs substantially increase and 
      results could be obstructed if all the biomarkers need to be tested with a single 
      cytometer. In this protocol, after collection and immediate incubation at 37  degrees C 
      for liquefication, semen samples were further analyzed for sperm apoptosis using 
      Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The 
      MMP was labeled with 
      5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-benzimidazolylcarbocyanine iodide 
      (JC-1) probe, and DNA damage was assessed using the sperm chromatin structure 
      assay (SCSA) with acridine orange (AO) staining. Thus, flow cytometric analysis 
      of sperm function markers can be a practical and reliable toolkit for the 
      diagnosis of infertility and evaluation of sperm function at both bench and bed.
FAU - Ling, Xi
AU  - Ling X
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Zou, Peng
AU  - Zou P
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Ao, Lin
AU  - Ao L
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Zhou, Niya
AU  - Zhou N
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Wang, Xiaogang
AU  - Wang X
AD  - Department of Chemical Defense Medicine, College of Military Preventive Medicine, 
      Army Medical University (Third Military Medical University).
FAU - Sun, Lei
AU  - Sun L
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Yang, Huan
AU  - Yang H
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Liu, Jinyi
AU  - Liu J
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Cao, Jia
AU  - Cao J
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University).
FAU - Chen, Qing
AU  - Chen Q
AD  - Institute of Toxicology, College of Preventive Medicine, Army Medical University 
      (Third Military Medical University); chenqingforward@gmail.com.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20220421
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Biomarkers)
RN  - 0 (Chromatin)
SB  - IM
MH  - Humans
MH  - Male
MH  - Biomarkers
MH  - Chromatin
MH  - Flow Cytometry/methods
MH  - *Infertility
MH  - *Spermatozoa
EDAT- 2022/05/10 06:00
MHDA- 2022/05/12 06:00
CRDT- 2022/05/09 09:04
PHST- 2022/05/09 09:04 [entrez]
PHST- 2022/05/10 06:00 [pubmed]
PHST- 2022/05/12 06:00 [medline]
AID - 10.3791/63790 [doi]
PST - epublish
SO  - J Vis Exp. 2022 Apr 21;(182). doi: 10.3791/63790.