Measurement of bacterial size via image analysis of epifluorescence preparations: Description of an inexpensive system and solutions to some of the most common problems

Scientia Marina

Volumn 61, Issue 3, 1997, Pages 397-407

  Massana, Ramon ^a   Gasol, Josep M ^a   Bjørnsen, Peter K ^b   Blackburn, Nicholas ^c   Hagström, Åke ^d   Hietanen, Susanna ^e   Hygum, Bent H ^d   Kuparinen, Jorma ^e   Pedrós Alió, Carlos ^a  

^a INSTITUT DE CIÈNCIES DEL MAR   (Spain)

^b UNIVERSITY OF COPENHAGEN   (Denmark)

^c UMEÅ UNIVERSITY   (Sweden)

^d NATIONAL ENVIRONMENTAL RESEARCH INSTITUTE   (Denmark)

^e FINNISH INSTITUTE OF MARINE RESEARCH   (Finland)

Author keywords

Bacterial size; Epifluorescence microscopy; Image analysis

Indexed keywords

BACTERIA (MICROORGANISMS);

ANALYTICAL TECHNIQUES; BACTERIA; EPIFLUORESCENCE MICROSCOPY; SIZE MEASUREMENT;

EID: 0031399514     PISSN: 02148358     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Article

References (35)

1. Alldredge, A.L., J.J. Cole and D.A. Caron. - 1986. Production of heterotrophic bacteria inhabiting macroscopic organic aggregates (marine snow) from surface waters. Limnol. Oceanogr. 31: 68-78.
2. Billen, G., P. Servais and S. Becquevort. - 1990. Dynamics of bacterioplankton in oligotrophic and eutrophic aquatic environments: bottom-up or top-down control? Hydrobiologia 207: 37-42.
3. Bjørnsen, P.K. - 1986. Automatic determination of bacterioplankton biomass by image analysis. Appl. Environ. Microbiol. 51: 1199-1204.
4. Bjørnsen, P.K., B. Riemann, J. Pock-Steen, T.G. Nielsen and S.J. Horsted. - 1989. Regulation of bacterioplankton production and cell volume in a eutrophic estuary. Appl. Environ. Microbiol. 55: 1512-1518.
5. Bjørnsen, P.K., Å. Hagström, B. Hygum, T.G. Nielsen, N. Blackburn, R. Massana, C. Pedrós-Alió, C. Svarer, L.K. Hansen, S. Hietanen and J. Kuparinen. - 1995. DIADEME: Digital Image Analysis Development in European Marine Ecology. Scientific Report of the Contract No. MAS2-CT93-0077.
6. Bloem, J., M. Veninga and J. Shepherd. - 1995. Fully automatic determination of soil bacterium numbers, cell volumes, and frequencies of dividing cells by confocal laser scanning microscopy and image analysis. Appl. Environ. Microbiol. 61: 926-936.
7. Chrzanowski, T.H., R.D. Crotty and G.J. Hubbard. - 1987. Seasonal variation in cell volume of epilimnetic bacteria. Microb. Ecol. 16: 155-163.
8. Cole, J.J., M.L. Pace, N.F. Caraco and G.S. Steinhart. - 1993. Bacterial biomass and cell size distributions in lakes: More and larger cells in anoxic waters. Limnol. Oceanogr. 38: 1627-1632.
9. del Giorgio, P.A., D.F. Bird, Y.T. Prairie and D. Planas. - 1996. The flow cytometric determination of bacterial abundance in lake plankton using the green nucleic acid stain SYTO 13. Limnol. Oceanogr. 41: 783-789.
10. Estep, K.W., F. MacIntyre, E. Hjörleifsson and J.McN. Sieburth. - 1986. MacImage: a user-friendly image-analysis system for the accurate mensuration of marine organisms. Mar. Ecol. Prog. Ser. 33: 243-253.
11. Fry, J.C. - 1990. Direct methods and biomass estimation. Meth. Microbiol. 22: 41-85.
12. Fuhrman, J.A. - 1981. Influence of methods on the apparent size distribution of bacterioplankton cells: epifluorescence microscopy compared to scanning electron microscopy. Mar. Ecol. Prog. Ser. 5: 103-106.
13. Gasol, J.M., P.A. del Giorgio, R. Massana and C.M. Duarte. - 1995. Active versus inactive bacteria: size-dependence in a coastal marine plankton community. Mar. Ecol. Progr. Ser. 128: 91-97.
14. Gasol, J.M., R. Massana and C. Pedrós-Alió. - in press. Bacterial size structure as a method to analyze communities. In: Proceedings of the Seventh International Symposium of Microbial Ecology, Brazilian Soc. Microbiol., Santos, Brazil.
15. Hygum, B.H. - 1995. Abundance, volume and distribution of bacterioplankton and sub-micron particles in the open Baltic Sea. Proc. Sixth Int. Work. Meas. Microb. Act. Cycl. Matter in Aquat. Envir., Konstanz, Germany.
16. Jürgens, K. and H. Güde. - 1994. The potential importance of grazing-resistant bacteria in planktonic systems. Mar. Ecol. Prog. Ser. 112: 169-188.
17. Krambeck, C., H.-J. Krambeck, D. Schröder and S.Y. Newell. - 1990. Sizing bacterioplankton: a juxtaposition of bias due to shrinkage, halos, subjectivity in image interpretation and asymmetric distributions. Binary 2: 5-14.
18. Lee, S.H., Y.-C. Kang and J.A. Fuhrman. - 1995. Imperfect retention of natural bacterioplankton cells by glass fiber filters. Mar. Ecol. Prog. Ser. 119: 285-290.
19. Li, W.K.W., J.F. Jellett and P.M. Dickie. - 1995. DNA distributions in planktonic bacteria stained with TOTO or TO-PRO. Limnol. Oceanogr. 40: 1485-1495.
20. Novitsky, J.A. and R.Y. Morita. - 1976. Morphological characterization of small cells resulting from nutrient starvation of a psychrophilic marine vibrio. Appl. Environ. Microbiol. 32: 617-622.
21. Peters, R.H. - 1983. The ecological implications of body size. Cambridge Univ. Press, Cambridge, Massachusetts, USA.
22. Porter, K.G. and Y.S. Feig. - 1980. The use of DAPI for identification and enumeration of aquatic microflora. Limnol. Oceanogr. 25: 943-948.
23. Psenner, R. - 1991. Determination of bacterial cell volumes by image analysis. Verh. Internat. verein. Limnol. 24: 2605-2608.
24. Psenner, R. - 1993. Determination of size and morphology of aquatic bacteria by automated image analysis. In: P.F. Kemp, B.F. Sherr, E.B. Sherr and J.J. Cole (Eds.): Handbook of Methods in Aquatic Microbial Ecology, pp: 339-345. Lewis Publ., Boca Raton, Florida, USA.
25. Robertson, B.R. and D.K. Button. - 1989. Characterizing aquatic bacteria according to population, cell size, and apparent DNA content by flow cytometry. Cytometry 10: 70-76.
26. Schröder, D. and H.-J. Krambeck. - 1991. Advances in digital image analysis of bacterioplankton with epifluorescence microscopy. Verh. Internat. verein. Limnol. 24: 2601-2604.
27. Sieracki, M.E., P.W. Johnson and J.McN. Sieburth. - 1985. Detection, enumeration and sizing of planktonic bacteria by image-analysed epifluorescence microscopy. Appl. Environ. Microbiol. 49: 799-810.
28. Sieracki, M.E., S.E. Reichenbach and K.L. Webb. - 1989a. Evaluation of automated threshold selection methods for accurately sizing microscopic fluorescent cells by image analysis. Appl. Environ. Microbiol. 55: 2762-2772.
29. Sieracki, M.E., C.L. Viles and K.L. Webb. - 1989b. Algorithm to estimate cell biovolume using image analyzed microscopy. Cytometry 10: 551-557.
30. Sieracki, M.E. and K.L. Webb. - 1991. The application of image-analyzed fluorescence microscopy for characterizing planktonic bacteria and protists. In: P.C. Reid, C.M. Turley and P.H. Burkill (Eds.): Protozoa and their role in marine processes, pp. 77-100. NATO ASI Ser. vol. 25, Springer Verlag, Berlin.
31. Sieracki, M.E., E.M. Haugen and T.L. Cucci. - 1995. Overestimation of heterotrophic bacteria in the Sargasso Sea: direct evidence by flow and imaging cytometry. Deep-Sea Res. 42: 1399-1409.
32. Sommaruga, R. and R. Psenner. - 1995. Permanent presence of grazing-resistant bacteria in a hypertrophic lake. Appl. Environ. Microbiol. 61: 3457-3459.
33. Verity, P.G. and M.E. Sieracki. - 1993. Use of color image analysis and epifluorescence microscopy to measure plankton biomass. In: P.F. Kemp, B.F. Sherr, E.B. Sherr and J.J. Cole (Eds.): Handbook of Methods in Aquatic Microbial Ecology, pp. 327-338. Lewis Publ., Boca Raton, Florida, USA.
34. Viles, C.L. and M.E. Sieracki. - 1992. Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy. Appl. Environ. Microbiol. 58: 584-592.
35. White, P.A., J. Kalff, J.B. Rasmussen and J.M. Gasol. - 1991. The effect of temperature and algal biomass on bacterial production and specific growth rate in freshwater and marine habitats. Microb. Ecol. 21: 99-118.