PMID- 20097826
OWN - NLM
STAT- MEDLINE
DCOM- 20100518
LR  - 20220318
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Print)
IS  - 0099-2240 (Linking)
VI  - 76
IP  - 6
DP  - 2010 Mar
TI  - Agreement, precision, and accuracy of epifluorescence microscopy methods for 
      enumeration of total bacterial numbers.
PG  - 1981-91
LID - 10.1128/AEM.01724-09 [doi]
AB  - To assess interchangeability of estimates of bacterial abundance by different 
      epifluorescence microscopy methods, total bacterial numbers (TBNs) determined by 
      most widely accepted protocols were statistically compared. Bacteria in a set of 
      distinctive samples were stained with acridine orange (AO), 
      4'-6-diamidino-2-phenylindole (DAPI), and BacLight and enumerated by visual 
      counting (VC) and supervised image analysis (IA). Model II regression and 
      Bland-Altman analysis proved general agreements between IA and VC methods, 
      although IA counts tended to be lower than VC counts by 7% on a logarithmic 
      scale. Distributions of cells and latex beads on polycarbonate filters were best 
      fitted to negative binomial models rather than to Poisson or log-normal models. 
      The fitted models revealed higher precisions of TBNs by the IA method than those 
      by the VC method. In pairwise comparisons of the staining methods, TBNs by AO and 
      BacLight staining showed good agreement with each other, but DAPI staining had 
      tendencies of underestimation. Although precisions of the three staining methods 
      were comparable to one another (intraclass correlation coefficients, 0.97 to 
      0.98), accuracy of the DAPI staining method was rebutted by disproportionateness 
      of TBNs between pairs of samples that carried 2-fold different volumes of 
      identical cell suspensions. It was concluded that the TBN values estimated by AO 
      and BacLight staining are relatively accurate and interchangeable for 
      quantitative interpretation and that IA provides better precision than does VC. 
      As a prudent measure, it is suggested to avoid use of DAPI staining for 
      comparative studies investigating accuracy of novel cell-counting methods.
FAU - Seo, Eun-Young
AU  - Seo EY
AD  - Departement of Environmental Science, Kangwon National University, Chuncheon 
      200-701, Republic of Korea.
FAU - Ahn, Tae-Seok
AU  - Ahn TS
FAU - Zo, Young-Gun
AU  - Zo YG
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100122
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Fluorescent Dyes)
SB  - IM
MH  - Bacteria/*isolation & purification
MH  - Colony Count, Microbial/methods
MH  - Fluorescent Dyes/pharmacology
MH  - Image Processing, Computer-Assisted/methods
MH  - Microscopy, Fluorescence/methods
MH  - Staining and Labeling/methods
MH  - Statistics as Topic
PMC - PMC2837990
EDAT- 2010/01/26 06:00
MHDA- 2010/05/19 06:00
PMCR- 2010/09/01
CRDT- 2010/01/26 06:00
PHST- 2010/01/26 06:00 [entrez]
PHST- 2010/01/26 06:00 [pubmed]
PHST- 2010/05/19 06:00 [medline]
PHST- 2010/09/01 00:00 [pmc-release]
AID - AEM.01724-09 [pii]
AID - 1724-09 [pii]
AID - 10.1128/AEM.01724-09 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2010 Mar;76(6):1981-91. doi: 10.1128/AEM.01724-09. Epub 
      2010 Jan 22.