Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells
文献类型:期刊论文
作者 | Berry, David1; Mader, Esther1; Lee, Tae Kwon1; Woebken, Dagmar1; Wang, Yun3,4; Zhu, Di5; Palatinszky, Marton1; Schintimeister, Arno1,2; Schmid, Markus C.1; Hanson, Buck T.1 |
刊名 | PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA |
出版日期 | 2015-01-13 |
卷号 | 112期号:2页码:E194-E203 |
关键词 | ecophysiology single-cell microbiology carbohydrate utilization nitrifier Raman microspectroscopy |
英文摘要 | Microbial communities are essential to the function of virtually all ecosystems and eukaryotes, including humans. However, it is still a major challenge to identify microbial cells active under natural conditions in complex systems. In this study, we developed a new method to identify and sort active microbes on the single-cell level in complex samples using stable isotope probing with heavy water (D2O) combined with Raman microspectroscopy. Incorporation of D2O-derived D into the biomass of autotrophic and heterotrophic bacteria and archaea could be unambiguously detected via C-D signature peaks in single-cell Raman spectra, and the obtained labeling pattern was confirmed by nanoscale-resolution secondary ion MS. In fast-growing Escherichia coli cells, label detection was already possible after 20 min. For functional analyses of microbial communities, the detection of D incorporation from D2O in individual microbial cells via Raman microspectroscopy can be directly combined with FISH for the identification of active microbes. Applying this approach to mouse cecal microbiota revealed that the host-compound foragers Akkermansia muciniphila and Bacteroides acidifaciens exhibited distinctive response patterns to amendments of mucin and sugars. By Raman-based cell sorting of active (deuterated) cells with optical tweezers and subsequent multiple displacement amplification and DNA sequencing, novel cecal microbes stimulated by mucin and/or glucosamine were identified, demonstrating the potential of the nondestructive D2O-Raman approach for targeted sorting of microbial cells with defined functional properties for single-cell genomics. |
WOS标题词 | Science & Technology |
类目[WOS] | Multidisciplinary Sciences |
研究领域[WOS] | Science & Technology - Other Topics |
关键词[WOS] | 16S RIBOSOMAL-RNA ; IN-SITU HYBRIDIZATION ; HYDROGEN ISOTOPE FRACTIONATION ; BACTEROIDES-THETAIOTAOMICRON ; SP-NOV. ; RAMAN MICROSPECTROSCOPY ; GEN.-NOV. ; AKKERMANSIA-MUCINIPHILA ; OLIGONUCLEOTIDE PROBES ; COMMUNITY STRUCTURE |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000347732300014 |
源URL | [http://ir.qibebt.ac.cn/handle/337004/6102] |
专题 | 青岛生物能源与过程研究所_单细胞中心 |
作者单位 | 1.Univ Vienna, Dept Microbiol & Ecosyst Sci, Div Microbial Ecol, A-1090 Vienna, Austria 2.Univ Vienna, Fac Life Sci, Large Instrument Facil Adv Isotope Res, A-1090 Vienna, Austria 3.Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Chinese Acad Sciences Key Lab Biofuels, Single Cell Ctr, Shandong 266101, Peoples R China 4.Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Shandong Key Lab Energy Genet, Shandong 266101, Peoples R China 5.Univ Sheffield, Kroto Res Inst, Sheffield S3 7QH, S Yorkshire, England 6.Univ Sheffield, Dept Chem, Sheffield S3 7QH, S Yorkshire, England 7.Agr Res Org, Inst Anim Sci, Dept Ruminant Sci, IL-50250 Bet Dagan, France 8.Univ Vienna, Max F Perutz Labs, Dept Microbiol Immunobiol & Genet, A-1030 Vienna, Austria 9.Univ Jena, Inst Phys Chem, D-07743 Jena, Germany 10.Univ Jena, Abbe Ctr Photon, D-07743 Jena, Germany |
推荐引用方式 GB/T 7714 | Berry, David,Mader, Esther,Lee, Tae Kwon,et al. Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2015,112(2):E194-E203. |
APA | Berry, David.,Mader, Esther.,Lee, Tae Kwon.,Woebken, Dagmar.,Wang, Yun.,...&Wagner, Michael.(2015).Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,112(2),E194-E203. |
MLA | Berry, David,et al."Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 112.2(2015):E194-E203. |
入库方式: OAI收割
来源:青岛生物能源与过程研究所
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