A simple and highly efficient fixation method for Chrysochromulina polylepis (Prymnesiophytes) for analytical flow...

Area of Science:

• Marine biology
• Phytoplankton research
• Flow cytometry applications

Background:

• Studying fragile Prymnesiophyte species like Chrysochromulina polylepis requires an effective fixation method for flow cytometry (FC).
• The fixation method must ensure high cell yield for efficient FC measurements and rapid sample processing.
• A key requirement is the ability to store fixed samples for later FC analysis.

Purpose of the Study:

• To identify an effective fixation method for Chrysochromulina polylepis for flow cytometry (FC) analysis.
• To optimize fixation for high cell recovery, signal quality, and sample storage.
• To enable efficient and timely FC analysis of this delicate phytoplankton species.

Main Methods:

• Testing various aldehydes and methanol concentrations for cell fixation.
• Employing gravity sedimentation and centrifugation to concentrate fixed cells.
• Evaluating storage conditions for fixed samples.

Main Results:

• 0.25% glutaraldehyde (GA) fixation achieved approximately 90% cell recovery.
• Excellent signals were obtained during FC analysis of GA-fixed cells.
• GA-fixed cells can be centrifuged and stored for several weeks without compromising analysis.

Conclusions:

• Glutaraldehyde (GA) is the recommended fixative for FC analysis of C. polylepis due to high recovery rates and signal quality.
• Centrifugation of GA-fixed cells increases concentration, reducing FC measurement times.
• The ability to store fixed cells allows for flexible scheduling of FC analysis.