PMID- 35926128
OWN - NLM
STAT- MEDLINE
DCOM- 20220808
LR  - 20220808
IS  - 2691-1299 (Electronic)
IS  - 2691-1299 (Linking)
VI  - 2
IP  - 8
DP  - 2022 Aug
TI  - Sperm Chromatin Structure Assay (SCSA(R)) for Fertility Assessment.
PG  - e508
LID - 10.1002/cpz1.508 [doi]
AB  - The Sperm Chromatin Structure Assay (SCSA((R)) ) is a federally registered protocol 
      for simultaneous flow cytometric measures of sperm DNA integrity and chromatin 
      structure. Fresh or frozen/thawed raw semen samples are diluted in buffer to a 
      sperm concentration of  approximately 1-2x10(6) /ml and then treated with a pH 1.20 buffer for 
      30 s to open the DNA strands at sites of DNA strand breaks. The sperm are then 
      stained with acridine orange (AO) that intercalates into double-strand DNA and 
      fluoresces green (515-530 BP filter) and stacks on single-strand DNA that 
      fluoresces red (630 LP filter) upon excitation from a 488 nm laser. The extent of 
      single and double DNA strand breaks (DNA fragmentation index, %DFI) and level of 
      excess nuclear histones (high DNA stainable sperm, %HDS) are simultaneously 
      measured in individual sperm. From the time a fresh or frozen/thawed semen sample 
      is received at the site of a flow cytometer (FCM) programmed for the SCSA 
      protocol, data can be obtained within about 10 min on 5-10x10(3) sperm. The %DFI 
      and %HDS can be determined by computer-gated regions on the green versus red 
      cytogram. Alternatively, a determination is made by transforming the green versus 
      red cytogram to a total DNA stainability (red + green fluorescence) versus 
      red/red + green fluorescence cytogram from which a frequency histogram is 
      produced and the %DFI calculated from it. The clinical threshold for human 
      natural or IUI fertilization is 25% DFI at which point the ART lab should 
      consider moving to ICSI fertilization. The clinical threshold for HDS is also 
      25%; values above this level may result in early embryo death due to abnormal 
      gene readout caused by the abnormal tertiary structure of chromatin. Numerous 
      lifestyle and environmental factors cause sperm DNA fragmentation. Reactive 
      oxygen species (ROS) play a significant role in DNA breakage. (c) 2022 The Authors. 
      Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Sperm 
      Chromatin Structure Assay (SCSA(R)) Basic Protocol 2: SCSA data analysis: 
      Calculations of %DFI and %HDS of semen samples by one of two methods Support 
      Protocol 1: SCSA sample collection and shipping Support Protocol 2: Flow 
      cytometer set up Support Protocol 3: Selection and use of reference samples.
CI  - (c) 2022 The Authors. Current Protocols published by Wiley Periodicals LLC.
FAU - Evenson, Donald P
AU  - Evenson DP
AD  - SCSA Diagnostics, Brookings, South Dakota.
AD  - Department of OB/GYN, Sandford Medical School, University of South Dakota, Sioux 
      Falls, South Dakota.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Curr Protoc
JT  - Current protocols
JID - 101773894
RN  - 0 (Chromatin)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Chromatin
MH  - DNA
MH  - Fertility
MH  - Humans
MH  - Male
MH  - *Semen
MH  - *Spermatozoa
OTO - NOTNLM
OT  - DNA integrity
OT  - SCSA
OT  - flow cytometry
OT  - male fertility
OT  - pregnancy outcomes
OT  - sperm chromatin structure assay
EDAT- 2022/08/05 06:00
MHDA- 2022/08/09 06:00
CRDT- 2022/08/04 15:22
PHST- 2022/08/04 15:22 [entrez]
PHST- 2022/08/05 06:00 [pubmed]
PHST- 2022/08/09 06:00 [medline]
AID - 10.1002/cpz1.508 [doi]
PST - ppublish
SO  - Curr Protoc. 2022 Aug;2(8):e508. doi: 10.1002/cpz1.508.