PMID- 28678425
OWN - NLM
STAT- MEDLINE
DCOM- 20180523
LR  - 20220223
IS  - 1552-4930 (Electronic)
IS  - 1552-4922 (Linking)
VI  - 91
IP  - 9
DP  - 2017 Sep
TI  - Semi-automated scoring of triple-probe FISH in human sperm using confocal 
      microscopy.
PG  - 859-866
LID - 10.1002/cyto.a.23126 [doi]
AB  - Structural and numerical sperm chromosomal aberrations result from abnormal 
      meiosis and are directly linked to infertility. Any live births that arise from 
      aneuploid conceptuses can result in syndromes such as Kleinfelter, Turners, XYY 
      and Edwards. Multi-probe fluorescence in situ hybridization (FISH) is commonly 
      used to study sperm aneuploidy, however manual FISH scoring in sperm samples is 
      labor-intensive and introduces errors. Automated scoring methods are continuously 
      evolving. One challenging aspect for optimizing automated sperm FISH scoring has 
      been the overlap in excitation and emission of the fluorescent probes used to 
      enumerate the chromosomes of interest. Our objective was to demonstrate the 
      feasibility of combining confocal microscopy and spectral imaging with 
      high-throughput methods for accurately measuring sperm aneuploidy. Our approach 
      used confocal microscopy to analyze numerical chromosomal abnormalities in human 
      sperm using enhanced slide preparation and rigorous semi-automated scoring 
      methods. FISH for chromosomes X, Y, and 18 was conducted to determine sex 
      chromosome disomy in sperm nuclei. Application of online spectral linear unmixing 
      was used for effective separation of four fluorochromes while decreasing data 
      acquisition time. Semi-automated image processing, segmentation, classification, 
      and scoring were performed on 10 slides using custom image processing and 
      analysis software and results were compared with manual methods. No significant 
      differences in disomy frequencies were seen between the semi automated and manual 
      methods. Samples treated with pepsin were observed to have reduced background 
      autofluorescence and more uniform distribution of cells. These results 
      demonstrate that semi-automated methods using spectral imaging on a confocal 
      platform are a feasible approach for analyzing numerical chromosomal aberrations 
      in sperm, and are comparable to manual methods. (c) 2017 International Society for 
      Advancement of Cytometry.
CI  - (c) 2017 International Society for Advancement of Cytometry.
FAU - Branch, Francesca
AU  - Branch F
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
FAU - Nguyen, GiaLinh
AU  - Nguyen G
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
FAU - Porter, Nicholas
AU  - Porter N
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
FAU - Young, Heather A
AU  - Young HA
AD  - Department of Epidemiology and Biostatistics, Milken Institute School of Public 
      Health, George Washington University, Washington, DC.
FAU - Martenies, Sheena E
AU  - Martenies SE
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
FAU - McCray, Nathan
AU  - McCray N
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
FAU - Deloid, Glen
AU  - Deloid G
AD  - Department of Environmental Health Harvard School of Public Health, Boston, 
      Massachusetts.
FAU - Popratiloff, Anastas
AU  - Popratiloff A
AD  - Nanofabrication and Imaging Center, George Washington University, Washington, DC.
FAU - Perry, Melissa J
AU  - Perry MJ
AD  - Department of Environmental and Occupational Health, Milken Institute School of 
      Public Health, George Washington University, Washington, DC.
LA  - eng
GR  - S10 RR025565/RR/NCRR NIH HHS/United States
GR  - R01 ES017457/ES/NIEHS NIH HHS/United States
GR  - S10 OD010710/OD/NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20170705
PL  - United States
TA  - Cytometry A
JT  - Cytometry. Part A : the journal of the International Society for Analytical 
      Cytology
JID - 101235694
SB  - IM
MH  - Adult
MH  - Aneuploidy
MH  - Cell Nucleus/genetics
MH  - Chromosome Aberrations
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Microscopy, Confocal/methods
MH  - Middle Aged
MH  - Sex Chromosomes/*genetics
MH  - Spermatozoa/*physiology
OTO - NOTNLM
OT  - Fluorescence in situ hybridization
OT  - aneuploidy
OT  - chromosomal abnormality
OT  - confocal microscopy
OT  - human spermatozoa
OT  - sex chromosome disomy
OT  - spectral imaging
EDAT- 2017/07/06 06:00
MHDA- 2018/05/24 06:00
CRDT- 2017/07/06 06:00
PHST- 2016/08/26 00:00 [received]
PHST- 2017/04/06 00:00 [revised]
PHST- 2017/04/13 00:00 [accepted]
PHST- 2017/07/06 06:00 [pubmed]
PHST- 2018/05/24 06:00 [medline]
PHST- 2017/07/06 06:00 [entrez]
AID - 10.1002/cyto.a.23126 [doi]
PST - ppublish
SO  - Cytometry A. 2017 Sep;91(9):859-866. doi: 10.1002/cyto.a.23126. Epub 2017 Jul 5.