PMID- 15583358
OWN - NLM
STAT- MEDLINE
DCOM- 20050218
LR  - 20131121
IS  - 1099-8004 (Print)
IS  - 1099-8004 (Linking)
VI  - 6
IP  - 3
DP  - 2005 Jan
TI  - Comparison of a visual to a computer-assisted technique for detecting apoptosis.
PG  - 180-6
AB  - In many studies, fluorescent dyes (ethidium bromide [EB] and acridine orange 
      [AO]) are used to stain DNA to determine if nuclei are apoptotic. However, there 
      are numerous visual methods for counting these stained DNA that may lead to 
      inaccuracies Measuring apoptosis by the visual counting method may be imprecise 
      because of the variability of individuals' perception of color. Therefore, the 
      authors compared a visual method of counting chromatin for apoptosis with a 
      method relying on a computer program. They began counting chromatin using the 
      visual method, in which individuals identify the stained DNA using their own 
      visual perception. For comparison, they used a software-based counting method 
      (analySIS software) to determine the color (hue) of the stained DNA. Using the 
      numeric hue values from the software eliminates the variations in human color 
      perception. Intra and interrater reliability of the visual and computer-assisted 
      counting methods were evaluated with Spearman's. The authors found statistical 
      significance in the intrarater reliability (r = 1.0, P = 0.0001 for all chromatin 
      categories) and interrater reliability (r = 0.975, P = 0.005 for both readings) 
      when using the software program. No statistical significance was found for the 
      visual counting method, indicating inaccuracy between and within raters. Thus, 
      the computer-assisted counting method of identifying the damaged DNA is more 
      accurate and precise than the individual's visual perception of color. Based on 
      these data, apoptosis measurements using color staining with EB and AO should be 
      determined using hue values generated by a computer program and not by a 
      researcher's visual assessment.
FAU - Goodyear-Bruch, Caryl
AU  - Goodyear-Bruch C
AD  - School of Nursing, University of Kansas, School of Allied Health, 3901 Rainbow 
      Blvd., Kansas City, KS 66160-7504, USA. cgoodyearbruch@kumc.edu
FAU - Simon, Kaycee
AU  - Simon K
FAU - Hall, Sandra
AU  - Hall S
FAU - Mayo, Matthew S
AU  - Mayo MS
FAU - Pierce, Janet D
AU  - Pierce JD
LA  - eng
GR  - R01 NR05317-03/NR/NINR NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biol Res Nurs
JT  - Biological research for nursing
JID - 9815758
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Free Radicals)
RN  - EN464416SI (Ethidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - *Apoptosis
MH  - Ethidium
MH  - Fluorescent Dyes
MH  - Free Radicals
MH  - Image Cytometry/*methods/statistics & numerical data
MH  - Image Processing, Computer-Assisted/*methods/statistics & numerical data
MH  - Male
MH  - Microscopy, Fluorescence/*methods/statistics & numerical data
MH  - Observer Variation
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Software
EDAT- 2004/12/08 09:00
MHDA- 2005/02/19 09:00
CRDT- 2004/12/08 09:00
PHST- 2004/12/08 09:00 [pubmed]
PHST- 2005/02/19 09:00 [medline]
PHST- 2004/12/08 09:00 [entrez]
AID - 6/3/180 [pii]
AID - 10.1177/1099800404271869 [doi]
PST - ppublish
SO  - Biol Res Nurs. 2005 Jan;6(3):180-6. doi: 10.1177/1099800404271869.