PMID- 7796694
OWN - NLM
STAT- MEDLINE
DCOM- 19950803
LR  - 20131121
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 19
IP  - 4
DP  - 1995 Apr 1
TI  - Comparative sperm chromatin structure assay measurements on epiillumination and 
      orthogonal axes flow cytometers.
PG  - 295-303
AB  - The sperm chromatin structure assay (SCSA) measures the susceptibility of sperm 
      nuclear DNA to acid-induced denaturation in situ, and was developed on two Ortho 
      flow cytometers, an FC200 [Becton Dickinson Immunocytometry Systems (BDIS), 
      Westwood, MA] and a Cytofluorograf 30 (BDIS), both having orthogonal axes of 
      fluorochrome excitation, emission, and sample flow. Sperm cells are first treated 
      with a pH 1.4 buffer to denature DNA in situ and then stained with the 
      metachromatic dye acridine orange (AO). The metachromatic fluorescence measured 
      reflects relative amounts of denatured (red fluorescence) and native (green 
      fluorescence) DNA present per cell. The extent of DNA denaturation is quantified 
      by the calculated parameter alpha t [alpha t = red/(red+green) fluorescence]. 
      Alpha t variables important for correlations with fertility and toxicant-induced 
      chromatin damage include mean (X alpha t), standard deviation (SD alpha t), and 
      cells outside the main population (COMP alpha t). Mean green fluorescence 
      intensity is an important measure for DNA content and/or degree of sperm 
      chromatin condensation. This study showed that the SCSA can be successfully run 
      on two epiillumination-type instruments, an Ortho ICP22A (BDIS, San Jose, CA) and 
      Skatron Argus (Tranby, Norway), and two additional orthogonal axes instruments, a 
      Becton Dickinson FACScan (BDIS) and a Coulter Elite (Coulter Corporation, 
      Hialeah, FL). Epiillumination instruments produced a different fluorescence 
      distribution than orthogonal instruments, but the resulting alpha t values showed 
      strong conformity and interpretation of results was the same. SCSA values 
      obtained on the Coulter Elite were most similar to the Cytofluorograf 30; the 
      FACScan green fluorescence distribution was narrower and allowed resolution of 
      cell doublets.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Evenson, D
AU  - Evenson D
AD  - Olson Biochemistry Laboratories, South Dakota State University, Brookings 57007, 
      USA.
FAU - Jost, L
AU  - Jost L
FAU - Gandour, D
AU  - Gandour D
FAU - Rhodes, L
AU  - Rhodes L
FAU - Stanton, B
AU  - Stanton B
FAU - Clausen, O P
AU  - Clausen OP
FAU - De Angelis, P
AU  - De Angelis P
FAU - Coico, R
AU  - Coico R
FAU - Daley, A
AU  - Daley A
FAU - Becker, K
AU  - Becker K
AU  - et al.
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Chromatin)
RN  - 9007-49-2 (DNA)
RN  - AT5C31J09G (Methyl Methanesulfonate)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Cattle
MH  - Chromatin/drug effects/*ultrastructure
MH  - DNA/chemistry/drug effects
MH  - Flow Cytometry/*instrumentation
MH  - Horses
MH  - Humans
MH  - Male
MH  - Methyl Methanesulfonate/toxicity
MH  - Mice
MH  - Nucleic Acid Denaturation/drug effects
MH  - Sheep
MH  - Spermatozoa/drug effects/*ultrastructure
MH  - Swine
MH  - Turkeys
EDAT- 1995/04/01 00:00
MHDA- 1995/04/01 00:01
CRDT- 1995/04/01 00:00
PHST- 1995/04/01 00:00 [pubmed]
PHST- 1995/04/01 00:01 [medline]
PHST- 1995/04/01 00:00 [entrez]
AID - 10.1002/cyto.990190403 [doi]
PST - ppublish
SO  - Cytometry. 1995 Apr 1;19(4):295-303. doi: 10.1002/cyto.990190403.