PMID- 12583717
OWN - NLM
STAT- MEDLINE
DCOM- 20030428
LR  - 20061115
IS  - 0723-2020 (Print)
IS  - 0723-2020 (Linking)
VI  - 25
IP  - 4
DP  - 2002 Dec
TI  - Improvement of ciliate identification and quantification: a new protocol for 
      fluorescence in situ hybridization (FISH) in combination with silver stain 
      techniques.
PG  - 555-71
AB  - A new protocol for taxon specific probe based fluorescent in situ hybridization 
      was developed for the identification and quantification of ciliates in microbial 
      communities. Various fixatives and experimental parameters were evaluated and 
      optimized with respect to cell permeability and morphological preservation. 
      Optimum results were adaption by obatined of a modified fixation method using 
      Bouin's solution. Furthermore, conventional staining procedures such as different 
      Protargol stain techniques and a silver nitrate impregnation method were modified 
      and can now be applied in combination with fluorescence in situ hybridization. 
      The new protocol allows a rapid and reliable identification as well as 
      quantification of ciliates based upon classical morphological aspects and rRNA 
      based phylogenetic relationships performed in one experiment. Furthermore, a set 
      of specific probes targeting different regions of the 18S rRNA was designed for 
      Glaucoma scintillans Ehrenberg, 1830 and tested by applying this new approach of 
      combining in situ cell hybridization with conventional staining techniques.
FAU - Fried, Johannes
AU  - Fried J
AD  - Institut fur Zoologie und Limnologie, Leopold-Franzens-Universitat Innsbruck, 
      Innsbruck, Austria. johannes.fried@uibk.ac.at
FAU - Ludwig, Wolfgang
AU  - Ludwig W
FAU - Psenner, Roland
AU  - Psenner R
FAU - Schleifer, Karl Heinz
AU  - Schleifer KH
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Syst Appl Microbiol
JT  - Systematic and applied microbiology
JID - 8306133
RN  - 0 (Fixatives)
RN  - 0 (Oligonucleotide Probes)
RN  - 0 (RNA, Ribosomal, 18S)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Ciliophora/*classification/genetics/*isolation & purification
MH  - Fixatives/chemistry
MH  - Fluorescence
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Molecular Sequence Data
MH  - Oligonucleotide Probes
MH  - RNA, Ribosomal, 18S/genetics
MH  - Sequence Homology
MH  - Silver Staining/*methods
MH  - Tetrahymena/isolation & purification
MH  - Water Microbiology
MH  - Water Purification/methods
EDAT- 2003/02/14 04:00
MHDA- 2003/04/29 05:00
CRDT- 2003/02/14 04:00
PHST- 2003/02/14 04:00 [pubmed]
PHST- 2003/04/29 05:00 [medline]
PHST- 2003/02/14 04:00 [entrez]
AID - S0723-2020(04)70146-0 [pii]
AID - 10.1078/07232020260517706 [doi]
PST - ppublish
SO  - Syst Appl Microbiol. 2002 Dec;25(4):555-71. doi: 10.1078/07232020260517706.