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Immunology and Infection

الكمي المتزامنة الخلوي ومكونات خارج الخلية من الأغشية الحيوية

Published: December 10, 2013 doi: 10.3791/50639

Materials

Name Company Catalog Number Comments
Bacto Agar Becton, Dickinson and Company 214010
Bacto Todd Hewitt Broth Becton, Dickinson and Company 249240
Yeast Extract, Granulated EMD Millipore 1.03753.0500
Bacto Tryptone Becton, Dickinson and Company 211705
OmniPur Sucrose EMD Millipore 8510
Potassium Chloride, ACS reagent, 99.0-100.5% Sigma-Aldrich P3911-500G
Potassium Phosphate, monobasic, ≥99.0%, ACS reagent Sigma-Aldrich P0662-500G
Sodium Chloride Sigma-Aldrich S9888-500G
Sodium Phosphate, Monobasic, Monohydrate EMD Millipore SX0710-1
Tris(hydroxymethyl)aminomethane, 99.8+%, ACS reagent Sigma-Aldrich 252859-500G
Concanavalin A, Alexa Fluor 647 Conjugate Invitrogen C21421
Syto 9 Invitrogen S34854
Sypro Red Invitrogen S12012 or S6653
Biotène PBF Oral Rinse GlaxoSmithKline N/A
Listerine Total Care McNeil-PPC, Inc. N/A

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References

  1. Costerton, J. W., Stewart, P. S., Greenberg, E. P. Bacterial biofilms: a common cause of persistent infections. Science. 284, 1318-1322 (1999).
  2. Hall-Stoodley, L., Costerton, J. W., Stoodley, P. Bacterial biofilms: from the natural environment to infectious diseases. Nat. Rev. Microbiol. 2, 95-108 (2004).
  3. Yang, Y., Sreenivasan, P. K., Subramanyam, R., Cummins, D. Multiparameter assessments to determine the effects of sugars and antimicrobials on a polymicrobial oral biofilm. Appl. Environ. Microbiol. 72, 6734-6742 (2006).
  4. Klein, M. I., Xiao, J., Heydorn, A., Koo, H. An analytical tool-box for comprehensive biochemical, structural and transcriptome evaluation of oral biofilms mediated by mutans streptococci. J. Vis. Exp. , (2011).
  5. Conchello, J. A., Lichtman, J. W. Optical sectioning microscopy. Nat. Methods. 2, 920-931 (2005).
  6. Robinson, J. P. Principles of confocal microscopy. Methods Cell Biol. 63, 89-106 (2001).
  7. Neu, T. R., Kuhlicke, U., Lawrence, J. R. Assessment of fluorochromes for two-photon laser scanning microscopy of biofilms. Appl. Environ. Microbiol. 68, 901-909 (2002).
  8. Wood, S. R., et al. Architecture of intact natural human plaque biofilms studied by confocal laser scanning microscopy. J. Dental Res. 79, 21-27 (2000).
  9. Chen, M. Y., Lee, D. J., Tay, J. H., Show, K. Y. Staining of extracellular polymeric substances and cells in bioaggregates. Appl. Microbiol. Biotechnol. 75, 467-474 (2007).
  10. Zinchuk, V., Zinchuk, O., Okada, T. Quantitative colocalization analysis of multicolor confocal immunofluorescence microscopy images: pushing pixels to explore biological phenomena. Acta Histochem. Cytochem. 40, 101-111 (2007).
  11. Dunn, K. W., Kamocka, M. M., McDonald, J. H. A practical guide to evaluating colocalization in biological microscopy. Am. J. Physiol. Cell Physiol. 300, C723-C742 (2011).
  12. Beyenal, H., Donovan, C., Lewandowski, Z., Harkin, G. Three-dimensional biofilm structure quantification. J. Microbiol. Methods. 59, 395-413 (2004).
الكمي المتزامنة الخلوي ومكونات خارج الخلية من الأغشية الحيوية
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Cite this Article

Khajotia, S. S., Smart, K. H.,More

Khajotia, S. S., Smart, K. H., Pilula, M., Thompson, D. M. Concurrent Quantification of Cellular and Extracellular Components of Biofilms. J. Vis. Exp. (82), e50639, doi:10.3791/50639 (2013).

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