PMID- 21755095
OWN - NLM
STAT- MEDLINE
DCOM- 20111213
LR  - 20211020
IS  - 1473-0189 (Electronic)
IS  - 1473-0197 (Print)
IS  - 1473-0189 (Linking)
VI  - 11
IP  - 16
DP  - 2011 Aug 21
TI  - Microfluidic fluorescence in situ hybridization and flow cytometry (muFlowFISH).
PG  - 2673-9
LID - 10.1039/c1lc20151d [doi]
AB  - We describe an integrated microfluidic device (muFlowFISH) capable of performing 
      16S rRNA fluorescence in situ hybridization (FISH) followed by flow cytometric 
      detection for identifying bacteria in natural microbial communities. The device 
      was used for detection of species involved in bioremediation of Cr(vi) and other 
      metals in groundwater samples from a highly-contaminated environmental site 
      (Hanford, WA, USA). The muFlowFISH seamlessly integrates two components: a 
      hybridization chamber formed between two photopolymerized membranes, where cells 
      and probes are electrophoretically loaded, incubated and washed, and a downstream 
      cross structure for electrokinetically focusing cells into a single-file flow for 
      flow cytometry analysis. The device is capable of analyzing a wide variety of 
      bacteria including aerobic, facultative and anaerobic bacteria and was initially 
      tested and validated using cultured microbes, including Escherichia coli, as well 
      as two strains isolated from Hanford site: Desulfovibrio vulgaris strain RCH1, 
      and Pseudomonas sp.strain RCH2 that are involved in Cr(vi) reduction and 
      immobilization. Combined labeling and detection efficiencies of 74-97% were 
      observed in experiments with simple mixtures of cultured cells, confirming 
      specific labeling. Results obtained were in excellent agreement with those 
      obtained by conventional flow cytometry confirming the accuracy of muFlowFISH. 
      Finally, the device was used for analyzing water samples collected on different 
      dates from the Hanford site. We were able to monitor the numbers of Pseudomonas 
      sp. with only 100-200 cells loaded into the microchip. The muFlowFISH approach 
      provides an automated platform for quantitative detection of microbial cells from 
      complex samples, and is ideally suited for analysis of precious samples with low 
      cell numbers such as those found at extreme environmental niches, bioremediation 
      sites, and the human microbiome.
CI  - This journal is (c) The Royal Society of Chemistry 2011
FAU - Liu, Peng
AU  - Liu P
AD  - Biotechnology and Bioengineering Department, Sandia National Laboratories, PO Box 
      969, MS 9291, Livermore, CA 94550, USA.
FAU - Meagher, Robert J
AU  - Meagher RJ
FAU - Light, Yooli K
AU  - Light YK
FAU - Yilmaz, Suzan
AU  - Yilmaz S
FAU - Chakraborty, Romy
AU  - Chakraborty R
FAU - Arkin, Adam P
AU  - Arkin AP
FAU - Hazen, Terry C
AU  - Hazen TC
FAU - Singh, Anup K
AU  - Singh AK
LA  - eng
GR  - R01 DE020891/DE/NIDCR NIH HHS/United States
GR  - R01 DE020891-03/DE/NIDCR NIH HHS/United States
GR  - 5 R01 DE020891/DE/NIDCR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20110714
PL  - England
TA  - Lab Chip
JT  - Lab on a chip
JID - 101128948
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Bacteria/*chemistry
MH  - Flow Cytometry/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Microfluidic Analytical Techniques/*methods
MH  - RNA, Ribosomal, 16S/*analysis
MH  - *Water Microbiology
PMC - PMC3145043
MID - NIHMS307139
EDAT- 2011/07/15 06:00
MHDA- 2011/12/14 06:00
PMCR- 2012/08/21
CRDT- 2011/07/15 06:00
PHST- 2011/07/15 06:00 [entrez]
PHST- 2011/07/15 06:00 [pubmed]
PHST- 2011/12/14 06:00 [medline]
PHST- 2012/08/21 00:00 [pmc-release]
AID - 10.1039/c1lc20151d [doi]
PST - ppublish
SO  - Lab Chip. 2011 Aug 21;11(16):2673-9. doi: 10.1039/c1lc20151d. Epub 2011 Jul 14.