PMID- 15458552
OWN - NLM
STAT- MEDLINE
DCOM- 20050215
LR  - 20220311
IS  - 0303-4569 (Print)
IS  - 0303-4569 (Linking)
VI  - 36
IP  - 5
DP  - 2004 Oct
TI  - Evaluation of chromatin integrity in human sperm using acridine orange staining 
      with different fixatives and after cryopreservation.
PG  - 321-6
AB  - Staining of cells with acridine orange (AO) has been widely accepted as a 
      predictor of DNA damage in many cell types. Because of variability of protocols 
      used in previous studies, the AO staining technique has not been widely accepted 
      as a screening test to predict DNA damage in human sperm. In order to further 
      validate the use of AO staining, sperm were evaluated using numerous variations 
      in the staining protocol. This study also elucidated the effects of 
      cryopreservation on sperm DNA. Sperm fixation in Carnoy's solution showed 
      significantly (P < 0.05) more DNA damage (29.9 +/- 4.5%) than 2% glutaraldehyde 
      (14.4 +/- 2.1%), 4% paraformaldehyde (5.5 +/- 1.7%), no fixation (15.8 +/- 4.3%) 
      but did not differ from Diff Quik solution (19.2 +/- 5.8%). No difference was 
      observed for sperm DNA damage assessment using a 0.2 m (15.5 +/- 3.2%) or 0.3 m 
      (14.9 +/- 3.3%) concentration of Na(2)HPO(4).7H(2)O in the AO staining solution. 
      Frozen-thawed semen samples showed increased damage to sperm DNA under both 
      Carnoy's (fresh: 10.9 +/- 1.3%; frozen: 30.8 +/- 2.9%; P < 0.05) and Diff Quik 
      fixation (fresh: 6.2 +/- 0.8; frozen: 17.1 +/- 2.5%P < 0.05). Present data also 
      showed that spermatozoa from some individuals are more prone to DNA damage after 
      freezing and thawing procedures than others. In conclusion, Carnoy's fixative 
      provides a better predictive value for DNA damage to sperm using AO staining. 
      Additionally, cryopreservation increased damage to the sperm DNA.
FAU - Chohan, K R
AU  - Chohan KR
AD  - Andrology and IVF Laboratories, University of Utah School of Medicine, 675 
      Arapeen Drive, Suite 205, Salt Lake City, UT 84108, USA.
FAU - Griffin, J T
AU  - Griffin JT
FAU - Carrell, D T
AU  - Carrell DT
LA  - eng
PT  - Journal Article
PT  - Validation Study
PL  - Germany
TA  - Andrologia
JT  - Andrologia
JID - 0423506
RN  - 0 (Carnoy's solution)
RN  - 0 (Chromatin)
RN  - 0 (Fixatives)
RN  - 0 (Fluorescent Dyes)
RN  - 3K9958V90M (Ethanol)
RN  - 7V31YC746X (Chloroform)
RN  - F30N4O6XVV (Acridine Orange)
RN  - Q40Q9N063P (Acetic Acid)
SB  - IM
MH  - Acetic Acid/standards
MH  - *Acridine Orange
MH  - Chloroform/standards
MH  - Chromatin/*metabolism
MH  - *Cryopreservation
MH  - *DNA Damage
MH  - Ethanol/standards
MH  - Fixatives/standards
MH  - *Fluorescent Dyes
MH  - Humans
MH  - Male
MH  - Predictive Value of Tests
MH  - Semen Preservation/*adverse effects
MH  - Spermatozoa/*metabolism
MH  - Staining and Labeling
EDAT- 2004/10/02 05:00
MHDA- 2005/02/16 09:00
CRDT- 2004/10/02 05:00
PHST- 2004/10/02 05:00 [pubmed]
PHST- 2005/02/16 09:00 [medline]
PHST- 2004/10/02 05:00 [entrez]
AID - AND626 [pii]
AID - 10.1111/j.1439-0272.2004.00626.x [doi]
PST - ppublish
SO  - Andrologia. 2004 Oct;36(5):321-6. doi: 10.1111/j.1439-0272.2004.00626.x.