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Science China Chemistry

, Volume 61, Issue 7, pp 792–796 | Cite as

Antibiotics-based fluorescent probes for selective labeling of Gram-negative and Gram-positive bacteria in living microbiotas

  • Wei Wang
  • Xing Chen
Communications

Abstract

Gram-staining distinguishes bacteria into two major groups, Gram-positive and Gram-negative bacteria, and has become an essential technique in microbiology. However, Gram-staining is not compatible with living cells and thus limits its applications. Here, we report the development of a polymyxin B-based fluorescent probe that enables selective labeling of Gram-negative in the living microbiota samples. We first synthesized the polymyxin B-Cy3 conjugate and confirmed its specificity for labeling Gram-negative bacteria. In combination with a previously developed Gram-positive-specific fluorescent probe, we demonstrate two-color imaging of Gram-positive and Gram-negative bacteria in various kinds living microbiotas, including mouse gut, human oral, soil, and crude oil microbiotas, with high selectivity and coverage. Finally, a pilot use of the probes in staining bacteria on heat-fixed sputum smear was also demonstrated, showing its potentials in clinical microbiology. Our method provides a versatile tool for distinguishing Gram-positive and Gram-negative bacteria in both basic research and clinical settings.

Keywords

microbiota imaging Gram staining polymyxin B differential bacterial labeling 

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Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (21672013, 21425204, 21521003) and the National Key Research and Development Projects (2016YFA0501500). We thank Dr. Xiaolei Wu for providing the crude oil microbiota and the National Center for Protein Sciences Beijing (Peking University, Beijing, China) for help with flow cytometry analyses.

Supplementary material

11426_2018_9236_MOESM1_ESM.pdf (1.4 mb)
Polymyxin-based fluorescent probe for selective labeling of Gram-negative bacteria in living microbiotas

References

  1. 1.
    Beveridge T. Biotech Histochem, 2001, 76: 111–118CrossRefGoogle Scholar
  2. 2.
    Lozupone CA, Stombaugh JI, Gordon JI, Jansson JK, Knight R. Nature, 2012, 489: 220–230CrossRefGoogle Scholar
  3. 3.
    Forster S, Snape JR, Lappin-Scott HM, Porter J. Appl Environ Microbiol, 2002, 68: 4772–4779CrossRefGoogle Scholar
  4. 4.
    Holm C, Jespersen L. Appl Environ Microbiol, 2003, 69: 2857–2863CrossRefGoogle Scholar
  5. 5.
    McClelland R. Med Lab Obs, 2001, 33: 20–22Google Scholar
  6. 6.
    Gram C. Fortschr Med, 1884, 2: 185–189Google Scholar
  7. 7.
    Mason DJ, Shanmuganathan S, Mortimer FC, Gant VA. Appl Environ Microbiol, 1998, 64: 2681–2685Google Scholar
  8. 8.
    Clauß M, Springorum AC, Hartung J. Aerosol Sci Tech, 2012, 46: 818–827CrossRefGoogle Scholar
  9. 9.
    Sizemore RK, Caldwell JJ, Kendrick AS. Appl Environ Microbiol, 1990, 56: 2245–2247Google Scholar
  10. 10.
    Kajiwara H, Toda M, Mine T, Nakada H, Wariishi H, Yamamoto T. Microbes Environ, 2010, 25: 152–155CrossRefGoogle Scholar
  11. 11.
    Wang W, Zhu Y, Chen X. Biochemistry, 2017, 56: 3889–3893CrossRefGoogle Scholar
  12. 12.
    Trimble MJ, Mlynárčik P, Kolář M, Hancock REW. Cold Spring Harb Perspect Med, 2016, 6: a025288CrossRefGoogle Scholar
  13. 13.
    Abdelraouf K, Chang KT, Yin T, Hu M, Tam VH. Antimicrob Agents Chemother, 2014, 58: 4200–4202CrossRefGoogle Scholar
  14. 14.
    Schindler PRG, Teuber M. Antimicrob Agents Chemother, 1975, 8: 95–104CrossRefGoogle Scholar
  15. 15.
    Moore RA, Bates NC, Hancock RE. Antimicrob Agents Chemother, 1986, 29: 496–500CrossRefGoogle Scholar
  16. 16.
    Deris ZZ, Swarbrick JD, Roberts KD, Azad MAK, Akter J, Horne AS, Nation RL, Rogers KL, Thompson PE, Velkov T, Li J. Bioconjugate Chem, 2014, 25: 750–760CrossRefGoogle Scholar
  17. 17.
    Moison E, Xie R, Zhang G, Lebar MD, Meredith TC, Kahne D. ACS Chem Biol, 2017, 12: 928–932CrossRefGoogle Scholar
  18. 18.
    Soon RL, Velkov T, Chiu F, Thompson PE, Kancharla R, Roberts K, Larson I, Nation RL, Li J. Anal Biochem, 2011, 409: 273–283CrossRefGoogle Scholar
  19. 19.
    Azad MAK, Yun B, Roberts KD, Nation RL, Thompson PE, Velkov T, Li J. Antimicrob Agents Chemother, 2014, 58: 6337–6338CrossRefGoogle Scholar
  20. 20.
    Zavascki AP, Goldani LZ, Li J, Nation RL. J Antimicrob Chemother, 2007, 60: 1206–1215CrossRefGoogle Scholar
  21. 21.
    Hancock R. Trends Microbiol, 1997, 5: 37–42CrossRefGoogle Scholar
  22. 22.
    Fierer N, Bradford MA, Jackson RB. Ecology, 2007, 88: 1354–1364CrossRefGoogle Scholar
  23. 23.
    Cai M, Nie Y, Chi CQ, Tang YQ, Li Y, Wang XB, Liu ZS, Yang Y, Zhou J, Wu XL. Sci Rep, 2015, 5: 16057CrossRefGoogle Scholar

Copyright information

© Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Molecular Medicine, Renji Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  2. 2.College of Chemistry and Molecular Engineering, Beijing National Laboratory for Molecular Sciences, Peking-Tsinghua Center for Life Sciences, Synthetic and Functional Biomolecules Center, and Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of EducationPeking UniversityBeijingChina

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