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PMID: 16534939 Published · ppublish English Journal Article

Bacterial growth on surfaces: automated image analysis for quantification of growth rate-related parameters.

Applied and environmental microbiology ·Vol. 61 ·No. 2 ·1995-02-00 ·Pages 741-8

Moller S, Kristensen CS, Poulsen LK, Carstensen JM, Molin S

Abstract

A fast routine method for estimating bacterial cell growth rates by using the metachromatic dye acridine orange is described. The method allows simultaneous estimates of cellular RNA and DNA contents of single cells. Acridine orange staining can be used as a nonspecific supplement to quantitative species-specific hybridizations with fluorescence-labelled ribosomal probes to estimate the single-cell concentration of RNA. By automated analysis of digitized images of stained cells, we determined four independent growth rate-related parameters: cellular RNA and DNA contents, cell volume, and the frequency of dividing cells in a cell population. These parameters were used to compare physiological states of liquid-suspended and surface-growing Pseudomonas putida KT2442 in chemostat cultures. The major finding is that the correlation between substrate availability and cellular growth rate found for the free-living cells was not observed for the surface-bound cells; in contrast, the data indicate an almost constant growth rate for attached cells which was independent of the dilution rate in the chemostat.

Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Moller S
Kristensen C S
Poulsen L K
Carstensen J M
Molin S
References (47)
47 references, click to expand
  1. Frequency of dividing cells, a new approach to the determination of bacterial growth rates in aquatic environments.
    Appl Environ Microbiol. 1979 May;37(5):805-12 PMID: 16345378
  2. Morphological analysis of nuclear separation and cell division during the life cycle of Escherichia coli.
    J Bacteriol. 1976 Jan;125(1):248-57 PMID: 1107308
  3. Effect of interfaces on small, starved marine bacteria.
    Appl Environ Microbiol. 1982 May;43(5):1166-72 PMID: 16346013
  4. Automatic determination of bacterioplankton biomass by image analysis.
    Appl Environ Microbiol. 1986 Jun;51(6):1199-204 PMID: 16347077
  5. Measurement of Glucose Utilization by Pseudomonas fluorescens That Are Free-Living and That Are Attached to Surfaces.
    Appl Environ Microbiol. 1986 Oct;52(4):672-6 PMID: 16347163
  6. Estimating the growth rate of slowly growing marine bacteria from RNA content.
    Appl Environ Microbiol. 1993 Aug;59(8):2594-601 PMID: 16349018
  7. The Effect of Solid Surfaces upon Bacterial Activity.
    J Bacteriol. 1943 Jul;46(1):39-56 PMID: 16560677
  8. Use of rRNA fluorescence in situ hybridization for measuring the activity of single cells in young and established biofilms.
    Appl Environ Microbiol. 1993 May;59(5):1354-60 PMID: 7685999
  9. Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system.
    J Histochem Cytochem. 1977 Jan;25(1):46-56 PMID: 64567
  10. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology.
    Appl Environ Microbiol. 1988 May;54(5):1079-84 PMID: 3389805
  11. Microfluorometric characterization of intracellular nucleic acids and nucleoproteins by acridine orange.
    Acta Physiol Scand Suppl. 1966;267:1-122 PMID: 4161117
  12. Sorption of heterotrophic and enteric bacteria to glass surfaces in the continuous culture of river water.
    Appl Microbiol. 1974 Oct;28(4):572-8 PMID: 4424694
  13. The conformation of the ribonucleic acid in ribosomes. Dye stacking studies.
    Biochemistry. 1966 Sep;5(9):3044-56 PMID: 5336402
  14. Ribonucleic acid composition of bacteria as a function of growth rate.
    J Mol Biol. 1966 Jul;18(2):308-20 PMID: 5338757
  15. Flow cytometry of bacteria: a promising tool in experimental and clinical microbiology.
    J Gen Microbiol. 1983 Apr;129(4):973-80 PMID: 6350534
  16. Simultaneous determination of the total number of aquatic bacteria and the number thereof involved in respiration.
    Appl Environ Microbiol. 1978 Dec;36(6):926-35 PMID: 367268
  17. The concentrations of stable RNA and ribosomes in Rickettsia prowazekii.
    Mol Microbiol. 1994 Apr;12(1):115-20 PMID: 7520114
  18. Molecular and microscopic identification of sulfate-reducing bacteria in multispecies biofilms.
    Appl Environ Microbiol. 1992 Feb;58(2):614-23 PMID: 1376982
  19. Ribosomes exist in large excess over the apparent demand for protein synthesis during carbon starvation in marine Vibrio sp. strain CCUG 15956.
    J Bacteriol. 1992 Nov;174(21):6780-8 PMID: 1383195
  20. Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy.
    Appl Environ Microbiol. 1992 Feb;58(2):584-92 PMID: 1610183
  21. Use of a fluorescent redox probe for direct visualization of actively respiring bacteria.
    Appl Environ Microbiol. 1992 Jun;58(6):1801-8 PMID: 1622256
  22. Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology.
    J Bacteriol. 1990 Feb;172(2):762-70 PMID: 1688842
  23. The differential fluorescence of bacteria stained with acridine orange and the effects of heat.
    J Appl Bacteriol. 1991 Jul;71(1):51-8 PMID: 1716625
  24. Optical sectioning of microbial biofilms.
    J Bacteriol. 1991 Oct;173(20):6558-67 PMID: 1917879
  25. Use of oligodeoxynucleotide signature probes for identification of physiological groups of methylotrophic bacteria.
    Appl Environ Microbiol. 1990 Sep;56(9):2858-65 PMID: 1967116
  26. Influence of interfaces on microbial activity.
    Microbiol Rev. 1990 Mar;54(1):75-87 PMID: 2181260
  27. Bacterial evolution.
    Microbiol Rev. 1987 Jun;51(2):221-71 PMID: 2439888
  28. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells.
    J Bacteriol. 1988 Feb;170(2):720-6 PMID: 2448289
  29. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells.
    Science. 1989 Mar 10;243(4896):1360-3 PMID: 2466341
  30. Evaluation of automated threshold selection methods for accurately sizing microscopic fluorescent cells by image analysis.
    Appl Environ Microbiol. 1989 Nov;55(11):2762-72 PMID: 2516431
  31. Algorithm to estimate cell biovolume using image analyzed microscopy.
    Cytometry. 1989 Sep;10(5):551-7 PMID: 2776571
  32. Bacterial biofilms in nature and disease.
    Annu Rev Microbiol. 1987;41:435-64 PMID: 3318676
  33. Adsorption and adhesion processes in microbial growth at interfaces.
    Adv Colloid Interface Sci. 1986 Jun;25(1):59-86 PMID: 3333130
  34. Biofilms, the customized microniche.
    J Bacteriol. 1994 Apr;176(8):2137-42 PMID: 8157581
  35. Identification of coccoid Escherichia coli BJ4 cells in the large intestine of streptomycin-treated mice.
    Infect Immun. 1993 Dec;61(12):5029-34 PMID: 8225579
  36. Responses to nutrient starvation in Pseudomonas putida KT2442: analysis of general cross-protection, cell shape, and macromolecular content.
    J Bacteriol. 1994 Jan;176(1):7-14 PMID: 8282712
  37. In situ identification of bacteria in drinking water and adjoining biofilms by hybridization with 16S and 23S rRNA-directed fluorescent oligonucleotide probes.
    Appl Environ Microbiol. 1993 Jul;59(7):2293-8 PMID: 8357261
  38. Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes.
    Gene. 1993 Jan 15;123(1):17-24 PMID: 8380783
  39. Survival of nonculturable Aeromonas salmonicida in lake water.
    Appl Environ Microbiol. 1993 Mar;59(3):874-80 PMID: 8481011
  40. Acridine orange staining reaction as an index of physiological activity in Escherichia coli.
    J Microbiol Methods. 1991;13:87-97 PMID: 11543591
  41. Dependency on medium and temperature of cell size and chemical composition during balanced grown of Salmonella typhimurium.
    J Gen Microbiol. 1958 Dec;19(3):592-606 PMID: 13611202
  42. Studies on the role of ribonucleic acid in the growth of bacteria.
    Biochim Biophys Acta. 1960 Jul 29;42:99-116 PMID: 13728193
  43. Relation between mitotic index, duration of mitosis, generation time and fraction of dividing cells in a cell population.
    Nature. 1962 Feb 10;193:555-6 PMID: 13914174
  44. A microscope-based flow cytophotometer.
    Histochem J. 1983 Feb;15(2):147-60 PMID: 6853222
  45. Distribution of sulfate-reducing bacteria, O2, and H2S in photosynthetic biofilms determined by oligonucleotide probes and microelectrodes.
    Appl Environ Microbiol. 1993 Nov;59(11):3840-9 PMID: 7506896
  46. Quantification of methanogenic groups in anaerobic biological reactors by oligonucleotide probe hybridization.
    Appl Environ Microbiol. 1994 Apr;60(4):1241-8 PMID: 7517129
  47. Microcomputer-assisted biomass determination of plankton bacteria on scanning electron micrographs.
    Appl Environ Microbiol. 1981 Jul;42(1):142-9 PMID: 16345807
Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.61.2.741-748.1995
Published
1995-02-00
Pages
741-8
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC1388357
External Links
PubMed source DOI: 10.1128/aem.61.2.741-748.1995 PMC Full Text View journal details
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