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Optical plankton analyser: a flow cytometer for plankton analysis, I: Design considerations
J C Peeters1, G B Dubelaar, J Ringelberg
1Tidal Waters Division, Rijkswaterstaat, The Hague, The Netherlands.
Cytometry
|September 1, 1989
Summary
A new flow cytometer (FCM) design addresses limitations in analyzing diverse phytoplankton field samples. This instrument accurately measures a wide range of cell sizes and lengths at high flow rates, crucial for ecological studies.
Area of Science:
- Marine Biology
- Analytical Chemistry
- Instrumentation Engineering
Background:
- Standard flow cytometers (FCMs) struggle with the wide particle size distribution (0.5–500 µm diameter, >2000 µm length) and low concentrations typical of phytoplankton field samples.
- Existing FCMs face limitations including inadequate flow rates (minimum 4 µL/s), uneven illumination, incomplete processing of long signals, and mechanical stress causing fragmentation of filamentous colonies.
Purpose of the Study:
- To describe the design criteria for a novel flow cytometer (FCM) specifically engineered for analyzing field samples of phytoplankton.
- To overcome the limitations of commercially available FCMs in accurately measuring the broad range of phytoplankton sizes and morphologies at required flow rates.
Main Methods:
- Developed a new FCM design incorporating a slit-shaped focal spot, logarithmic amplifiers, and a pulse integration system capable of processing long signals to achieve a 5–6 decade dynamic range.
- Integrated multi-laser capabilities for species identification, a low fluid shear cuvette to prevent colony fragmentation, and a trigger gate extension for handling inhomogeneously fluorescent algal filaments.
Main Results:
- The new FCM design accommodates particle sizes from 0.5 to 500 µm in diameter and over 2000 µm in length.
- Achieved a dynamic range of 5 to 6 decades, enabling accurate analysis of low-concentration, diverse phytoplankton populations.
- The design minimizes fragmentation of filamentous colonies and ensures complete signal processing for accurate phytoplankton analysis.
Conclusions:
- The developed flow cytometer design effectively addresses the challenges of analyzing diverse phytoplankton field samples, offering improved accuracy and reliability over standard instruments.
- This specialized FCM is crucial for advancing phytoplankton research, enabling more precise ecological studies and monitoring of marine environments.
- The design innovations provide a robust platform for high-throughput analysis of phytoplankton, supporting a deeper understanding of marine microbial ecosystems.