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Characterization of Aquatic Biofilms with Flow Cytometry
Published on: June 6, 2018
Viable cell sorting of dinoflagellates by multiparametric flow cytometry
Christopher D Sinigalliano1, Jamie Winshell, Maria A Guerrero
1Southeast Environmental Research Center, Florida International University, Miami, FL 33199, USA.
Phycologia
|March 23, 2010
Summary
Electronic cell sorting offers a viable alternative to manual cell picking for isolating marine phytoplankton, including fragile dinoflagellates. While manual methods yield larger cultures, electronic sorting reduces labor and ensures culture purity, accelerating algal strain discovery.
Area of Science:
- Marine Biology
- Phytoplankton Ecology
- Cell Biology
Background:
- Traditional isolation of marine phytoplankton, such as dinoflagellates, relies on labor-intensive manual cell picking.
- Electronic cell sorting (ECS) presents a potential automated alternative for phytoplankton isolation and culture.
- Assessing the viability and efficiency of ECS compared to manual methods is crucial for advancing phytoplankton research.
Purpose of the Study:
- To compare the efficacy of electronic cell sorting (ECS) versus manual micropipette picking for isolating and culturing marine eukaryotic phytoplankton.
- To evaluate cell viability, culture success rates, and genetic diversity achieved by both isolation methods.
- To explore the potential of ECS in accelerating the discovery of new algal strains.
Main Methods:
- Comparison of ECS and manual cell picking for isolating dinoflagellates and other marine phytoplankton.
- Assessment of cell trauma and viability post-sorting using fragile species like Karenia brevis.
- Evaluation of large-scale culture success rates and the need for re-isolation.
- Application of broad and restrictive flow cytometric gating strategies.
- Testing of a novel recovery medium (RE) for axenic dinoflagellate cultures.
Main Results:
- ECS did not cause significant trauma to fragile dinoflagellates, yielding viable cells.
- Manual picking had a higher success rate for large-scale cultures (2%) compared to ECS (0.5%), but required more labor and re-isolation.
- ECS cultures required no re-isolation, ensuring unialgal status.
- Similar percentages of unique genotypes were obtained from both methods (57% manual vs. 54% ECS).
- A novel recovery medium (RE) achieved a 70% recovery rate for isolated cells.
- Direct ECS was more successful than pre-enrichment sorting followed by ECS.
Conclusions:
- Electronic cell sorting is a viable, less labor-intensive method for isolating marine phytoplankton, ensuring culture purity.
- While manual picking currently excels in large-scale culture initiation, ECS shows promise for accelerating algal strain discovery, especially when combined with automated screening.
- The development of optimized recovery media, like RE, is critical for enhancing ECS efficiency.
