Year: 2026
Type: preprint
Abstract: Microcystis spp. are difficult to pellet by centrifugation because of intracellular gas vesicles that make cells buoyant. The efficiency of pelleting is further influenced by culture conditions (batch vs. continuous culture) and the physiological state of cells. This variability complicates protocols that require reliable cell recovery, especially when maintaining the physiological state of cells is important. Previous studies have shown that disrupting the gas vesicles by exposing cells to high amounts of pressure (0.5 - 0.7 MPa) improves pelleting efficiency (Abeynayaka et al., 2016; Huang et al., 2017). However, this method involves the use of equipment that not all labs have access to and can be disruptive to cellular physiology. Therefore, a more accessible approach that minimizes physiological disturbances is needed. The method described in this protocol concentrates cells by filtration followed by washing cells off the filter with 1x phosphate-buffered saline (PBS), which has shown to consistently produce pellets after centrifugation. Using this approach, cell recovery averaged 99.5%, providing a reliable method for pelleting Microcystis spp. without the need to apply high amounts of pressure to collapse gas vesicles. This has been tested on M. aeruginosa PCC 7806 and ΔmcyB in both batch and continuous cultures.
Language: English
Subfield: Environmental Chemistry
Field: Environmental Science
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W7172436563