Detergent-based separation of microbes from marine particles

Area of Science:

• Marine microbiology
• Biogeochemistry
• Single-cell analysis

Background:

• Particle-associated microbes are crucial for marine biogeochemical cycles.
• Investigating these microbes requires separating them from marine particles.
• Existing methods for microbial dissociation from particles are understudied.

Purpose of the Study:

• To compare the effectiveness of Tween 20 and Tween 80 in dissociating particle-associated marine microbes.
• To evaluate the impact of these detergents on cell viability and community composition.
• To optimize a method for gentle microbial detachment from marine particles.

Main Methods:

• Comparison of Tween 20 and Tween 80 for microbial dissociation.
• Flow cytometry to quantify cell liberation and viability.
• 16S rRNA gene amplicon sequencing to analyze microbial community composition.
• Evaluation across different seasons and locations.

Main Results:

• Both Tween 20 and Tween 80 effectively dissociated microbes from particles with minimal cell mortality.
• Tween 80 liberated a higher number of particle-associated cells into the free-living fraction.
• Tween 80 treatments resulted in more uniform dissociated communities and enriched abundant members.
• Tween 80 proved most effective for gentle dissociation.

Conclusions:

• Tween 80 is recommended for effectively and gently dissociating particle-associated marine microbes.
• This optimized method facilitates downstream single-cell and molecular analyses.
• Understanding particle-associated microbial communities is vital for marine ecology.