Abstract

A novel cytometer is reported for measuring particle-size distribution, concentration, and biomass of marine phytoplankton containing chlorophyll a. The system utilizes optical fibers to carry light to and from a flow tube for measuring phytoplankton taken directly from the ocean. A unique feature of this system is in the simple optical detection scheme for which sample handling and preparation are not required. This simplicity makes the system especially suitable for field measurements. The system utilizes sophisticated digital signal processors to handle and reduce the large amount of data gathered. The signal-processing algorithms are vigorously streamlined to process the signals in real time, thus computing size and flow velocity information instead of logging the raw data. The high efficiency of the signal processors gives the system a performance throughput of ∼250 particles/s. The system was tested both in the laboratory and in the field, yielding good discrimination of size distribution and sensitivity of concentration.

© 2001 Optical Society of America

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References

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  1. G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
    [CrossRef] [PubMed]
  2. J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
    [CrossRef] [PubMed]
  3. A. Desiderio, C. Moore, C. Lantz, T. J. Cowles, “Multiple excitation fluorometer for in situ oceanographic applications,” Appl. Opt. 36, 1289–1296 (1997).
    [CrossRef] [PubMed]
  4. S. G. Ackleson, R. W. Spinrad, C. M. Yentsch, J. F. Brown, W. Korjeff-Bellows, “Phytoplankton optical properties: flow cytometric examinations of dilution-/induced effects,” Appl. Opt. 27, 1262–1269 (1988).
    [CrossRef] [PubMed]
  5. L. E. Drain, The Laser Doppler Technique (Wiley, New York, 1980).
  6. A. Radbruch, Flow Cytometry and Cell Sorting (Springer-Verlag, Berlin, 1992).
    [CrossRef]
  7. Y. Yeh, H. Z. Cummins, “Localized fluid flow measurements with a He–Ne laser spectrometer,” Appl. Phys. Lett. 4, 176–178 (1964).
    [CrossRef]
  8. P. G. Falkowski, Z. Kolber, “Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans,” Aust. J. Plant Physiol. 22, 341–335 (1995).
    [CrossRef]
  9. U. Schreiber, A. Krieger, “Two fundamentally different types of variable chlorophyll in vivo,” FEBS Lett. 397, 131–135 (1996).
    [CrossRef] [PubMed]

1999

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

1997

1996

U. Schreiber, A. Krieger, “Two fundamentally different types of variable chlorophyll in vivo,” FEBS Lett. 397, 131–135 (1996).
[CrossRef] [PubMed]

1995

P. G. Falkowski, Z. Kolber, “Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans,” Aust. J. Plant Physiol. 22, 341–335 (1995).
[CrossRef]

1988

1964

Y. Yeh, H. Z. Cummins, “Localized fluid flow measurements with a He–Ne laser spectrometer,” Appl. Phys. Lett. 4, 176–178 (1964).
[CrossRef]

Ackleson, S. G.

Baethmann, A.

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

Beeker, A. E. R.

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

Brown, J. F.

Cowles, T. J.

Cummins, H. Z.

Y. Yeh, H. Z. Cummins, “Localized fluid flow measurements with a He–Ne laser spectrometer,” Appl. Phys. Lett. 4, 176–178 (1964).
[CrossRef]

Desiderio, A.

Drain, L. E.

L. E. Drain, The Laser Doppler Technique (Wiley, New York, 1980).

Dubelaar, G. B. J.

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

Falkowski, P. G.

P. G. Falkowski, Z. Kolber, “Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans,” Aust. J. Plant Physiol. 22, 341–335 (1995).
[CrossRef]

Gerritzen, P. L.

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

Jonker, R. R.

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

Kachel, V.

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

Kolber, Z.

P. G. Falkowski, Z. Kolber, “Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans,” Aust. J. Plant Physiol. 22, 341–335 (1995).
[CrossRef]

Korjeff-Bellows, W.

Krieger, A.

U. Schreiber, A. Krieger, “Two fundamentally different types of variable chlorophyll in vivo,” FEBS Lett. 397, 131–135 (1996).
[CrossRef] [PubMed]

Lantz, C.

Moore, C.

Plesnila, N.

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

Radbruch, A.

A. Radbruch, Flow Cytometry and Cell Sorting (Springer-Verlag, Berlin, 1992).
[CrossRef]

Schreiber, U.

U. Schreiber, A. Krieger, “Two fundamentally different types of variable chlorophyll in vivo,” FEBS Lett. 397, 131–135 (1996).
[CrossRef] [PubMed]

Spinrad, R. W.

Tangen, K.

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

Wietzorrek, J.

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

Yeh, Y.

Y. Yeh, H. Z. Cummins, “Localized fluid flow measurements with a He–Ne laser spectrometer,” Appl. Phys. Lett. 4, 176–178 (1964).
[CrossRef]

Yentsch, C. M.

Appl. Opt.

Appl. Phys. Lett.

Y. Yeh, H. Z. Cummins, “Localized fluid flow measurements with a He–Ne laser spectrometer,” Appl. Phys. Lett. 4, 176–178 (1964).
[CrossRef]

Aust. J. Plant Physiol.

P. G. Falkowski, Z. Kolber, “Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans,” Aust. J. Plant Physiol. 22, 341–335 (1995).
[CrossRef]

Cytometry

G. B. J. Dubelaar, P. L. Gerritzen, A. E. R. Beeker, R. R. Jonker, K. Tangen, “Design and first results of CytoBuoy: a wireless flow cytometer for in-situ analysis of marine and fresh waters,” Cytometry 37, 247–254 (1999).
[CrossRef] [PubMed]

J. Wietzorrek, N. Plesnila, A. Baethmann, V. Kachel, “A new multiparameter flow cytometer: optical and electrical cell analysis in combination with video microscopy in flow,” Cytometry 35, 291–301 (1999).
[CrossRef] [PubMed]

FEBS Lett.

U. Schreiber, A. Krieger, “Two fundamentally different types of variable chlorophyll in vivo,” FEBS Lett. 397, 131–135 (1996).
[CrossRef] [PubMed]

Other

L. E. Drain, The Laser Doppler Technique (Wiley, New York, 1980).

A. Radbruch, Flow Cytometry and Cell Sorting (Springer-Verlag, Berlin, 1992).
[CrossRef]

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Figures (11)

Fig. 1
Fig. 1

Optical probe head setup: BS, beam splitter; PMT, photomultiplier tube.

Fig. 2
Fig. 2

Exaggerated angle of bow-tie measurement volume.

Fig. 3
Fig. 3

CASPA system.

Fig. 4
Fig. 4

Particle flowing through the center of the measurement volume.

Fig. 5
Fig. 5

Size distribution of Chlorella (a) from microscopic measurement, (b) from the cytometer, and (c) from the cytometer with approximately three times the concentration of (b).

Fig. 6
Fig. 6

Synchronous Doppler burst with LIF signal.

Fig. 7
Fig. 7

Waveforms showing the possible events: (a) two phytoplankton particles traversing through the measurement volume, (b) a phytoplankton particle following a detritus particle. LDV, laser Doppler velocimetry.

Fig. 8
Fig. 8

Size distributions of photoplankton at (a) Pak Sha Wan, (b) Wong Shek, (c) Tai Tam, and (d) Cafeteria Beach test sites.

Fig. 9
Fig. 9

Histograms of average phytoplankton concentrations at various sites.

Fig. 10
Fig. 10

Size distribution as a function of time.

Fig. 11
Fig. 11

Average photoplankton concentration as a function of time.

Equations (4)

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l=λ2n sinα/2,
f=2Vn cos β sinα/2λ,
f=2Vn sinα/2λ.
D=Vt-W.

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