A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy

  • J Microbiol Methods. 2012 Jun;89(3):216-21. doi: 10.1016/j.mimet.2012.03.005.
Elena Gorokhova  1 ,  Lisa Mattsson ,  Annica M Sundström
Affiliations
  • 1. Department of Applied Environmental Science, Stockholm University, SE-106 91 Stockholm, Sweden. [email protected]
PMID: 22446098 DOI: 10.1016/j.mimet.2012.03.005
Abstract

Two fluorescent dyes, TO-PRO-1 iodide and 5-CFDA-AM, were evaluated for LIVE/DEAD assessment of unicellular Marine algae Brachiomonas submarina and Tetraselmis suecica. Epifluorescence microscopy was used to estimate cell viability in predetermined mixtures of viable and non-viable algal cells and validated using microplate growth assay as reference measurements. On average, 5-CFDA-AM underestimated live cell abundance by ~25% compared with viability estimated by the growth assay, whereas TO-PRO-1 iodide provided accurate viability estimates. Furthermore, viability estimates based on staining with TO-PRO-1 iodide were not affected by a storage period of up to one month in -80°C, making the assay a good candidate for routine assessment of phytoplankton populations in field and laboratory studies.

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