PMID- 21507466
OWN - NLM
STAT- MEDLINE
DCOM- 20120516
LR  - 20220129
IS  - 1618-0372 (Electronic)
IS  - 0065-1281 (Print)
IS  - 0065-1281 (Linking)
VI  - 114
IP  - 2
DP  - 2012 Feb
TI  - Simultaneous 16S and 18S rRNA fluorescence in situ hybridization (FISH) on LR 
      White sections demonstrated in Vestimentifera (Siboglinidae) tubeworms.
PG  - 122-30
LID - 10.1016/j.acthis.2011.03.008 [doi]
AB  - Traditional morphological identification of invertebrate marine species is 
      limited in early life history stages for many taxa. In this study, we 
      demonstrate, by example of Vestimentiferan tubeworms (Siboglinidae, Polychaeta), 
      that the simultaneous fluorescence in situ hybridization (FISH) of both 
      eukaryotic host and bacterial symbiont cells is possible on a single semi-thin (1 
      mum) section. This allows the identification of host specimens to species level as 
      well as offering visualization of bacteria distributed within the host tissue. 
      Previously published 18S rRNA host-specific oligonucleotide probes for Riftia 
      pachyptila, Tevnia jerichonana and a newly designed Oasisia alvinae probe, as 
      well as a 16S rRNA probe targeting symbionts found in all host species, were 
      applied. A number of standard fixation and hybridization parameters were tested 
      and optimized for the best possible signal intensity and cellular resolution. 
      Ethanol conserved samples embedded in LR White low viscosity resin yielded the 
      best results with regard to both signal intensity and resolution. We show that 
      extended storage times of specimens does not affect the quality of signals 
      attained by FISH and use our protocol to identify morphologically unidentifiable 
      tubeworm individuals from a small data set, conforming to previous findings in 
      succession studies of the Siboglinidae family.
CI  - Copyright (c) 2011 Elsevier GmbH. All rights reserved.
FAU - Schimak, Mario P
AU  - Schimak MP
AD  - University of Vienna, Department of Marine Biology, Austria. 
      mario.schimak@gmail.com
FAU - Toenshoff, Elena R
AU  - Toenshoff ER
FAU - Bright, Monika
AU  - Bright M
LA  - eng
GR  - P 20282/FWF_/Austrian Science Fund FWF/Austria
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110419
PL  - Germany
TA  - Acta Histochem
JT  - Acta histochemica
JID - 0370320
RN  - 0 (Acrylic Resins)
RN  - 0 (DNA Probes)
RN  - 0 (Fixatives)
RN  - 0 (Polymers)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - 1HG84L3525 (Formaldehyde)
RN  - 94188-59-7 (LR white)
RN  - Y19UC83H8E (paraform)
SB  - IM
MH  - *Acrylic Resins
MH  - Animals
MH  - Base Sequence
MH  - Cryoultramicrotomy
MH  - DNA Probes/chemistry
MH  - Fixatives/chemistry
MH  - Formaldehyde/chemistry
MH  - Gammaproteobacteria/genetics/metabolism
MH  - In Situ Hybridization, Fluorescence
MH  - Molecular Typing/methods
MH  - *Plastic Embedding
MH  - Polychaeta/*genetics/metabolism/microbiology
MH  - Polymers/chemistry
MH  - RNA, Ribosomal, 16S/genetics/*metabolism
MH  - RNA, Ribosomal, 18S/genetics/*metabolism
MH  - Symbiosis
PMC - PMC3278570
EDAT- 2011/04/22 06:00
MHDA- 2012/05/17 06:00
PMCR- 2012/02/01
CRDT- 2011/04/22 06:00
PHST- 2011/02/07 00:00 [received]
PHST- 2011/03/18 00:00 [revised]
PHST- 2011/03/20 00:00 [accepted]
PHST- 2011/04/22 06:00 [entrez]
PHST- 2011/04/22 06:00 [pubmed]
PHST- 2012/05/17 06:00 [medline]
PHST- 2012/02/01 00:00 [pmc-release]
AID - S0065-1281(11)00056-0 [pii]
AID - 10.1016/j.acthis.2011.03.008 [doi]
PST - ppublish
SO  - Acta Histochem. 2012 Feb;114(2):122-30. doi: 10.1016/j.acthis.2011.03.008. Epub 
      2011 Apr 19.