PMID- 19492327
OWN - NLM
STAT- MEDLINE
DCOM- 20090914
LR  - 20090629
IS  - 1860-7314 (Electronic)
IS  - 1860-6768 (Linking)
VI  - 4
IP  - 6
DP  - 2009 Jun
TI  - Flow-through imaging cytometry for characterization of Salmonella subpopulations 
      in alfalfa sprouts, a complex food system.
PG  - 880-7
LID - 10.1002/biot.200800360 [doi]
AB  - We recently developed an approach combining fluorescence in situ hybridization 
      (FISH) and flow cytometry for detecting low levels of Salmonella spp. 
      (approximately 10(3) cells/mL sprout wash) against high levels of naturally 
      occurring sprout flora (approximately 10(7)-10(8) CFU/g sprouts). Although this 
      "FISH and flow" approach provided rapid presence/absence testing for Salmonella 
      in this complex food system, it was not capable of more nuanced tasks, such as 
      probing the phenotypic complexity of the microbes present in sprouts or 
      determining the physical interactions of Salmonella with these microbes, or with 
      sprout debris. In the present study, we have combined rapid FISH-based labeling 
      of Salmonella spp. in sprout washes with flow-through imaging cytometry (FT-IC), 
      using the ImageStream 100, a commercial FT-IC instrument. This approach enables 
      image-based characterization of various subpopulations of interest occurring 
      within these samples. Here, we demonstrate the ability of FT-IC to unambiguously 
      identify cells, cell aggregates and other events within these subpopulations 
      based on both cell morphology and hybridization status after reaction with a 
      Salmonella-targeted probe cocktail. Our ability to directly explore the nature of 
      these events expands the layers of information possible from cytometric analyses 
      of these complex samples and clearly demonstrates that "a picture is worth a 
      thousand dots".
FAU - Bisha, Bledar
AU  - Bisha B
AD  - Rapid Microbial Detection and Control Laboratory, Department of Food Science and 
      Human Nutrition, Iowa State University, Ames, IA 50011, USA.
FAU - Brehm-Stecher, Byron F
AU  - Brehm-Stecher BF
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Germany
TA  - Biotechnol J
JT  - Biotechnology journal
JID - 101265833
SB  - IM
MH  - Flow Cytometry/*methods
MH  - *Food Microbiology
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Medicago sativa/*microbiology
MH  - Microscopy, Electron, Scanning
MH  - Plant Shoots/microbiology
MH  - Salmonella typhimurium/*genetics
EDAT- 2009/06/06 09:00
MHDA- 2009/09/15 06:00
CRDT- 2009/06/04 09:00
PHST- 2009/06/04 09:00 [entrez]
PHST- 2009/06/06 09:00 [pubmed]
PHST- 2009/09/15 06:00 [medline]
AID - 10.1002/biot.200800360 [doi]
PST - ppublish
SO  - Biotechnol J. 2009 Jun;4(6):880-7. doi: 10.1002/biot.200800360.