Best practices in the flow cytometry of microalgae. Issue 4 (2nd March 2021)

• Record Type: Journal Article
• Authors: Čertnerová, Dora; Galbraith, David W.
• Other Names: Galbraith David guestEditor.
• Is Part Of: Cytometry. Volume 99:Issue 4(2021)
• Page Start: 359
• Page End: 364
• Publication Date: 2021-03-02
• Subjects: best practices -- microalgae -- flow cytometry -- nuclear isolation -- genome size -- algal FCM standards; Flow cytometry -- Periodicals; Imaging systems in biology -- Periodicals; Imaging systems in medicine -- Periodicals; Diagnostic imaging -- Periodicals; 571.605
• DOI: 10.1002/cyto.a.24328
• Languages: English
• ISSNs: 1552-4922

Abstract

Abstract: Microalgae are photosynthetic microorganisms with a major influence on global ecosystems. Further, owing to the production of various secondary metabolites, microalgae are also intensively studied for their enormous potential in biotechnology and its applications. While flow cytometry (FCM) is a fast and reliable method particularly suitable for genome size estimation in plant and animal studies, its application to microalgae often comes with many methodological challenges due to specific issues (e.g., cell wall composition, and presence of various secondary metabolites). Sample preparation requires considerable amounts of biomass, chemical fixation, and/or extraction of cellular components. In genome size estimation, appropriate methods for isolation of intact nuclei (using lysis buffers, razor‐blade chopping, various enzymes, or bead‐beating of cells) are essential for successful and high‐quality analyses. Nuclear DNA amounts of microalgae diverge greatly, varying by almost 30, 000‐fold (0.01 to 286 pg). Even though new algal reference standards for genome size are now being introduced, animal red blood cells and nuclei from plant tissues are still predominantly used. Due to our limited knowledge of microalgal life cycles, particular caution should be taken during 1C/2C‐value (or ploidy level) assignments.