PMID- 12596248
OWN - NLM
STAT- MEDLINE
DCOM- 20040210
LR  - 20220408
IS  - 1552-4922 (Print)
IS  - 1552-4922 (Linking)
VI  - 52
IP  - 1
DP  - 2003 Mar
TI  - Toluidine blue test for sperm DNA integrity and elaboration of image cytometry 
      algorithm.
PG  - 19-27
AB  - BACKGROUND: Sperm DNA integrity is of paramount importance in the prognosis of 
      fertility. We applied image cytometry to a toluidine blue (TB) test we recently 
      proposed. METHODS: Sperm samples from 33 men were assayed for standard sperm 
      parameters and classified as normal or abnormal. Sperm smears were subjected to 
      the TB test, DNA denaturation testing with acridine orange (AO), and terminal 
      deoxyuridine triphosphate biotin nick end labeling (TUNEL). In CCD image 
      analysis, TB-stained sperm cell heads were microscopically assigned to one of 
      four color groups (dark, blue, light violet, and light blue). The optical 
      densities of 6,600 cells in green and red CCD images were used to elaborate an 
      algorithm for discrimination of these groups. RESULTS: The proportions of sperm 
      in TB color groups, as estimated with the developed image cytometry algorithm, 
      correlated with microscopic features. The number of TB dark cells correlated with 
      the number of AO-red and TUNEL(+) cells. The proportion of TB dark cells in 
      normal samples did not exceed 35%. Light-blue sperm cell heads prevailed in 
      normal samples, whereas dark and blue sperm cell heads dominated in abnormal 
      samples. CONCLUSIONS: The TB test was suitable for the assessment of sperm cell 
      DNA integrity. The elaborated image cytometry algorithm can be used for this 
      purpose and for finer determination of sperm nucleus status.
CI  - Copyright 2003 Wiley-Liss, Inc.
FAU - Erenpreisa, Jekaterina
AU  - Erenpreisa J
AD  - Biomedical Research and Study Centre, University of Latvia, Riga, Latvia. 
      katrina@biomed.lu.lv
FAU - Erenpreiss, Juris
AU  - Erenpreiss J
FAU - Freivalds, Talivaldis
AU  - Freivalds T
FAU - Slaidina, Maija
AU  - Slaidina M
FAU - Krampe, Rasma
AU  - Krampe R
FAU - Butikova, Jelena
AU  - Butikova J
FAU - Ivanov, Andrey
AU  - Ivanov A
FAU - Pjanova, Dace
AU  - Pjanova D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry A
JT  - Cytometry. Part A : the journal of the International Society for Analytical 
      Cytology
JID - 101235694
RN  - 0 (Chromatin)
RN  - 15XUH0X66N (Tolonium Chloride)
RN  - 9007-49-2 (DNA)
RN  - EC 3.1.- (Deoxyribonucleases)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/pharmacology
MH  - Algorithms
MH  - Chromatin/metabolism
MH  - DNA/*metabolism
MH  - Deoxyribonucleases/chemistry
MH  - Humans
MH  - Image Cytometry/*methods
MH  - In Situ Nick-End Labeling
MH  - Male
MH  - Microscopy/*methods
MH  - Microscopy, Video
MH  - Spermatozoa/*metabolism
MH  - Tolonium Chloride/*pharmacology
EDAT- 2003/02/22 04:00
MHDA- 2004/02/11 05:00
CRDT- 2003/02/22 04:00
PHST- 2003/02/22 04:00 [pubmed]
PHST- 2004/02/11 05:00 [medline]
PHST- 2003/02/22 04:00 [entrez]
AID - 10.1002/cyto.a.10015 [doi]
PST - ppublish
SO  - Cytometry A. 2003 Mar;52(1):19-27. doi: 10.1002/cyto.a.10015.