PMID- 26475101
OWN - NLM
STAT- MEDLINE
DCOM- 20161006
LR  - 20240324
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Print)
IS  - 0099-2240 (Linking)
VI  - 82
IP  - 1
DP  - 2016 Jan 1
TI  - MiL-FISH: Multilabeled Oligonucleotides for Fluorescence In Situ Hybridization 
      Improve Visualization of Bacterial Cells.
PG  - 62-70
LID - 10.1128/AEM.02776-15 [doi]
AB  - Fluorescence in situ hybridization (FISH) has become a vital tool for 
      environmental and medical microbiology and is commonly used for the 
      identification, localization, and isolation of defined microbial taxa. However, 
      fluorescence signal strength is often a limiting factor for targeting all members 
      in a microbial community. Here, we present the application of a multilabeled FISH 
      approach (MiL-FISH) that (i) enables the simultaneous targeting of up to seven 
      microbial groups using combinatorial labeling of a single oligonucleotide probe, 
      (ii) is applicable for the isolation of unfixed environmental microorganisms via 
      fluorescence-activated cell sorting (FACS), and (iii) improves signal and imaging 
      quality of tissue sections in acrylic resin for precise localization of 
      individual microbial cells. We show the ability of MiL-FISH to distinguish 
      between seven microbial groups using a mock community of marine organisms and its 
      applicability for the localization of bacteria associated with animal tissue and 
      their isolation from host tissues using FACS. To further increase the number of 
      potential target organisms, a streamlined combinatorial labeling and spectral 
      imaging-FISH (CLASI-FISH) concept with MiL-FISH probes is presented here. Through 
      the combination of increased probe signal, the possibility of targeting 
      hard-to-detect taxa and isolating these from an environmental sample, the 
      identification and precise localization of microbiota in host tissues, and the 
      simultaneous multilabeling of up to seven microbial groups, we show here that 
      MiL-FISH is a multifaceted alternative to standard monolabeled FISH that can be 
      used for a wide range of biological and medical applications.
CI  - Copyright (c) 2015 Schimak et al.
FAU - Schimak, Mario P
AU  - Schimak MP
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany 
      mschimak@mpi-bremen.de bfuchs@mpi-bremen.de.
FAU - Kleiner, Manuel
AU  - Kleiner M
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany Energy 
      Bioengineering and Geomicrobiology Research Group, University of Calgary, 
      Calgary, Alberta, Canada.
FAU - Wetzel, Silke
AU  - Wetzel S
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany.
FAU - Liebeke, Manuel
AU  - Liebeke M
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany.
FAU - Dubilier, Nicole
AU  - Dubilier N
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany.
FAU - Fuchs, Bernhard M
AU  - Fuchs BM
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany 
      mschimak@mpi-bremen.de bfuchs@mpi-bremen.de.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151016
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Oligonucleotide Probes)
SB  - IM
MH  - Bacteria/cytology/*genetics
MH  - Flow Cytometry
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Oligonucleotide Probes/chemistry/*genetics
MH  - Staining and Labeling
PMC - PMC4702640
EDAT- 2015/10/18 06:00
MHDA- 2016/10/08 06:00
PMCR- 2015/12/22
CRDT- 2015/10/18 06:00
PHST- 2015/08/27 00:00 [received]
PHST- 2015/10/04 00:00 [accepted]
PHST- 2015/10/18 06:00 [entrez]
PHST- 2015/10/18 06:00 [pubmed]
PHST- 2016/10/08 06:00 [medline]
PHST- 2015/12/22 00:00 [pmc-release]
AID - AEM.02776-15 [pii]
AID - 02776-15 [pii]
AID - 10.1128/AEM.02776-15 [doi]
PST - epublish
SO  - Appl Environ Microbiol. 2015 Oct 16;82(1):62-70. doi: 10.1128/AEM.02776-15. Print 
      2016 Jan 1.