PMID- 24702905
OWN - NLM
STAT- MEDLINE
DCOM- 20150105
LR  - 20140520
IS  - 1618-0984 (Electronic)
IS  - 0723-2020 (Linking)
VI  - 37
IP  - 4
DP  - 2014 Jun
TI  - The effect of FISH and CARD-FISH on the isotopic composition of (13)C- and 
      (15)N-labeled Pseudomonas putida cells measured by nanoSIMS.
PG  - 267-76
LID - S0723-2020(14)00037-X [pii]
LID - 10.1016/j.syapm.2014.02.002 [doi]
AB  - The use of nanoSIMS for the exploration of microbial activities in natural 
      habitats often implies that stable isotope tracer experiments are combined with 
      in situ hybridization techniques (i.e. fluorescence in situ hybridization (FISH) 
      or catalyzed reporter deposition (CARD)-FISH). In this study, Pseudomonas putida 
      grown on (13)C- and (15)N-labeled carbon and nitrogen, collected in exponential 
      growth and stationary phases, was hybridized and analyzed by nanoSIMS. It was 
      shown that (13)C and (15)N fractions decreased after FISH and CARD-FISH in 
      comparison to chemically untreated cells. However, the fractions were influenced 
      differently by various treatments. After paraformaldehyde fixation of 
      exponentially growing cells, a reduction of the (13)C and (15)N fractions was 
      measured from 94+/-1.2% and 89.5+/-3.8% to 90.2+/-0.8% and 64+/-4.6%, respectively, 
      indicating that nitrogen isotopic composition was most influenced. A further 
      decrease of the (13)C and (15)N fractions to 80.7+/-6.5 and 59.5+/-4.1%, 
      respectively, was measured after FISH, while CARD-FISH decreased the fractions to 
      57.4+/-3.0% and 47.1+/-4.1%, respectively. The analysis of cells collected in 
      different growth phases revealed that the effect of various treatments seemed to 
      be dependent on the cell's physiological state. In addition, a mathematical model 
      that can be used in further studies was developed in order to calculate the 
      amount of carbon introduced into the cells by chemical treatments. These results 
      can be valuable for environmental FISH-nanoSIMS studies where the isotopic 
      composition of single cells will be used to quantitatively assess the importance 
      of specific populations to certain biochemical processes and determine budget 
      estimations.
CI  - Copyright (c) 2014 Elsevier GmbH. All rights reserved.
FAU - Musat, Niculina
AU  - Musat N
AD  - Helmholtz Centre for Environmental Research - UFZ, Department of Isotope 
      Biogeochemistry, Permoserstrasse 15, 04318 Leipzig, Germany. Electronic address: 
      niculina.musat@ufz.de.
FAU - Stryhanyuk, Hryhoriy
AU  - Stryhanyuk H
AD  - Helmholtz Centre for Environmental Research - UFZ, Department of Isotope 
      Biogeochemistry, Permoserstrasse 15, 04318 Leipzig, Germany.
FAU - Bombach, Petra
AU  - Bombach P
AD  - Helmholtz Centre for Environmental Research - UFZ, Department of Isotope 
      Biogeochemistry, Permoserstrasse 15, 04318 Leipzig, Germany.
FAU - Adrian, Lorenz
AU  - Adrian L
AD  - Helmholtz Centre for Environmental Research - UFZ, Department of Isotope 
      Biogeochemistry, Permoserstrasse 15, 04318 Leipzig, Germany.
FAU - Audinot, Jean-Nicolas
AU  - Audinot JN
AD  - Centre de Recherche Gabriel Lippmann, Department Science and Analysis of 
      Materials, 41, rue du Brill, L-4422 Belvaux, Luxembourg.
FAU - Richnow, Hans H
AU  - Richnow HH
AD  - Helmholtz Centre for Environmental Research - UFZ, Department of Isotope 
      Biogeochemistry, Permoserstrasse 15, 04318 Leipzig, Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140401
PL  - Germany
TA  - Syst Appl Microbiol
JT  - Systematic and applied microbiology
JID - 8306133
RN  - 0 (Carbon Isotopes)
RN  - 0 (Nitrogen Isotopes)
SB  - IM
MH  - Carbon Isotopes/metabolism
MH  - Environmental Microbiology
MH  - *In Situ Hybridization, Fluorescence
MH  - Isotope Labeling
MH  - Microbiological Techniques/methods
MH  - Models, Theoretical
MH  - Nitrogen Isotopes/metabolism
MH  - Pseudomonas putida/chemistry/*classification/*metabolism
MH  - Spectrometry, Mass, Secondary Ion/*methods
OTO - NOTNLM
OT  - CARD-FISH
OT  - FISH
OT  - NanoSIMS
OT  - Sample preparation
OT  - Single cell
OT  - Stable isotopes
EDAT- 2014/04/08 06:00
MHDA- 2015/01/06 06:00
CRDT- 2014/04/08 06:00
PHST- 2013/09/26 00:00 [received]
PHST- 2014/02/17 00:00 [revised]
PHST- 2014/02/18 00:00 [accepted]
PHST- 2014/04/08 06:00 [entrez]
PHST- 2014/04/08 06:00 [pubmed]
PHST- 2015/01/06 06:00 [medline]
AID - S0723-2020(14)00037-X [pii]
AID - 10.1016/j.syapm.2014.02.002 [doi]
PST - ppublish
SO  - Syst Appl Microbiol. 2014 Jun;37(4):267-76. doi: 10.1016/j.syapm.2014.02.002. 
      Epub 2014 Apr 1.