PMID- 37938730
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231110
IS  - 2730-6151 (Electronic)
IS  - 2730-6151 (Print)
IS  - 2730-6151 (Linking)
VI  - 2
IP  - 1
DP  - 2022 Jun 30
TI  - Correlative SIP-FISH-Raman-SEM-NanoSIMS links identity, morphology, biochemistry, 
      and physiology of environmental microbes.
PG  - 52
LID - 10.1038/s43705-022-00134-3 [doi]
LID - 52
AB  - Microscopic and spectroscopic techniques are commonly applied to study microbial 
      cells but are typically used on separate samples, resulting in population-level 
      datasets that are integrated across different cells with little spatial 
      resolution. To address this shortcoming, we developed a workflow that correlates 
      several microscopic and spectroscopic techniques to generate an in-depth analysis 
      of individual cells. By combining stable isotope probing (SIP), fluorescence in 
      situ hybridization (FISH), scanning electron microscopy (SEM), confocal Raman 
      microspectroscopy (Raman), and nano-scale secondary ion mass spectrometry 
      (NanoSIMS), we illustrate how individual cells can be thoroughly interrogated to 
      obtain information about their taxonomic identity, structure, physiology, and 
      metabolic activity. Analysis of an artificial microbial community demonstrated 
      that our correlative approach was able to resolve the activity of single cells 
      using heavy water SIP in conjunction with Raman and/or NanoSIMS and establish 
      their taxonomy and morphology using FISH and SEM. This workflow was then applied 
      to a sample of yet uncultured multicellular magnetotactic bacteria (MMB). In 
      addition to establishing their identity and activity, backscatter electron 
      microscopy (BSE), NanoSIMS, and energy-dispersive X-ray spectroscopy (EDS) were 
      employed to characterize the magnetosomes within the cells. By integrating these 
      techniques, we demonstrate a cohesive approach to thoroughly study environmental 
      microbes on a single-cell level.
CI  - (c) 2022. The Author(s).
FAU - Schaible, George A
AU  - Schaible GA
AD  - Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 
      59717, USA.
AD  - Center for Biofilm Engineering, Montana State University, Bozeman, MT, 59717, 
      USA.
AD  - Thermal Biology Institute, Montana State University, Bozeman, MT, 59717, USA.
FAU - Kohtz, Anthony J
AU  - Kohtz AJ
AD  - Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 
      59717, USA.
AD  - Center for Biofilm Engineering, Montana State University, Bozeman, MT, 59717, 
      USA.
AD  - Thermal Biology Institute, Montana State University, Bozeman, MT, 59717, USA.
FAU - Cliff, John
AU  - Cliff J
AUID- ORCID: 0000-0002-7395-5604
AD  - Environmental Molecular Sciences Laboratory, Pacific Northwest National 
      Laboratory, Richland, WA, 99354, USA.
FAU - Hatzenpichler, Roland
AU  - Hatzenpichler R
AUID- ORCID: 0000-0002-5489-3444
AD  - Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, 
      59717, USA. roland.hatzenpichler@montana.edu.
AD  - Center for Biofilm Engineering, Montana State University, Bozeman, MT, 59717, 
      USA. roland.hatzenpichler@montana.edu.
AD  - Thermal Biology Institute, Montana State University, Bozeman, MT, 59717, USA. 
      roland.hatzenpichler@montana.edu.
AD  - Department of Microbiology and Cell Biology, Montana State University, Bozeman, 
      MT, 59717, USA. roland.hatzenpichler@montana.edu.
LA  - eng
GR  - NNX17AK85G/National Aeronautics and Space Administration (NASA)/
GR  - 80NSSC20K1365/National Aeronautics and Space Administration (NASA)/
GR  - 80NSSC20K1365/National Aeronautics and Space Administration (NASA)/
GR  - 5999/Gordon and Betty Moore Foundation (Gordon E. and Betty I. Moore Foundation)/
GR  - DBI-1726561/National Science Foundation (NSF)/
GR  - SR-2017331/M.J. Murdock Charitable Trust (Murdock Charitable Trust)/
PT  - Journal Article
DEP - 20220630
PL  - England
TA  - ISME Commun
JT  - ISME communications
JID - 9918205372406676
PMC - PMC9723565
EDAT- 2022/06/30 00:00
MHDA- 2022/06/30 00:01
PMCR- 2022/06/30
CRDT- 2023/11/08 11:55
PHST- 2022/03/29 00:00 [received]
PHST- 2022/06/09 00:00 [accepted]
PHST- 2022/05/23 00:00 [revised]
PHST- 2022/06/30 00:01 [medline]
PHST- 2022/06/30 00:00 [pubmed]
PHST- 2023/11/08 11:55 [entrez]
PHST- 2022/06/30 00:00 [pmc-release]
AID - 10.1038/s43705-022-00134-3 [pii]
AID - 134 [pii]
AID - 10.1038/s43705-022-00134-3 [doi]
PST - epublish
SO  - ISME Commun. 2022 Jun 30;2(1):52. doi: 10.1038/s43705-022-00134-3.