PMID- 11823205
OWN - NLM
STAT- MEDLINE
DCOM- 20020328
LR  - 20210526
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 68
IP  - 2
DP  - 2002 Feb
TI  - Comparison of fluorescently labeled oligonucleotide and polynucleotide probes for 
      the detection of pelagic marine bacteria and archaea.
PG  - 661-7
AB  - We compared the detection of bacteria and archaea in the coastal North Sea and at 
      Monterey Bay, Calif., after fluorescence in situ hybridization (FISH) either with 
      rRNA-targeted oligonucleotide probes monolabeled with the cyanin dye Cy3 
      (oligoFISH) or with fluorescein-labeled polyribonucleotide probes (polyFISH). 
      During an annual cycle in German Bight surface waters, the percentages of 
      bacteria visualized by polyFISH (annual mean, 77% of total counts) were 
      significantly higher than those detected by oligoFISH (53%). The fraction of 
      total bacteria visualized by oligoFISH declined during winter, whereas cell 
      numbers determined by polyFISH remained constant throughout the year. Depth 
      profiles from Monterey Bay showed large differences in the fraction of bacterial 
      cells visualized by polyFISH and oligoFISH in the deeper water layers 
      irrespective of the season. Image-analyzed microscopy indicated that the superior 
      detection of cells by polyFISH with fluorescein-labeled probes in 
      bacterioplankton samples was less a consequence of higher absolute fluorescence 
      intensities but was rather related to quasi-linear bleaching dynamics and to a 
      higher signal-to-background ratio. The relative abundances of archaea in North 
      Sea and Monterey Bay spring samples as determined by oligoFISH were on average 
      higher than those determined by polyFISH. However, simultaneous hybridizations 
      with oligonucleotide probes for bacteria and archaea suggested that the oligoFISH 
      probe ARCH915 unspecifically stained a population of bacteria. Using either FISH 
      technique, blooms of archaea were observed in North Sea surface waters during the 
      spring and summer months. Marine group II archaea (Euryarchaeota) reached >30% of 
      total picoplankton abundances, as determined by polyFISH. We suggest that studies 
      of pelagic microbial community structure using oligoFISH with monolabeled probes 
      should focus on environments that yield detections > or =70% of total cell 
      counts, e.g., coastal surface waters during spring and summer.
FAU - Pernthaler, Annelie
AU  - Pernthaler A
AD  - Max-Planck-Institute for Marine Microbiology, Celsiusstrasse 1, D-28359 Bremen, 
      Germany.
FAU - Preston, Christina M
AU  - Preston CM
FAU - Pernthaler, Jakob
AU  - Pernthaler J
FAU - DeLong, Edward F
AU  - DeLong EF
FAU - Amann, Rudolf
AU  - Amann R
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Carbocyanines)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (Polyribonucleotides)
RN  - 0 (RNA, Ribosomal)
RN  - 0 (cyanine dye 3)
RN  - TPY09G7XIR (Fluorescein)
SB  - IM
MH  - Archaea/classification/genetics/*isolation & purification
MH  - Bacteria/classification/genetics/*isolation & purification
MH  - Carbocyanines/metabolism
MH  - Fluorescein/metabolism
MH  - Fluorescent Dyes/metabolism
MH  - Image Processing, Computer-Assisted
MH  - *In Situ Hybridization, Fluorescence
MH  - Oligonucleotide Probes/*genetics
MH  - Polyribonucleotides/*genetics
MH  - RNA, Ribosomal/genetics
MH  - Seawater/*microbiology
PMC - PMC126737
EDAT- 2002/02/02 10:00
MHDA- 2002/03/29 10:01
PMCR- 2002/02/01
CRDT- 2002/02/02 10:00
PHST- 2002/02/02 10:00 [pubmed]
PHST- 2002/03/29 10:01 [medline]
PHST- 2002/02/02 10:00 [entrez]
PHST- 2002/02/01 00:00 [pmc-release]
AID - 1668 [pii]
AID - 10.1128/AEM.68.2.661-667.2002 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2002 Feb;68(2):661-7. doi: 10.1128/AEM.68.2.661-667.2002.