Original paper

Live sorting and survival of unstained and DAPI-stained ciliates by flow cytometry

Lindström, Eva S.; Stadler, Peter; Weisse, Thomas

Archiv für Hydrobiologie Volume 157 Number 2 (2003), p. 173 - 184

published: Jun 11, 2003

DOI: 10.1127/0003-9136/2003/0157-0173

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ArtNo. ESP141015772002, Price: 29.00 €

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Abstract

The potential of flow cytometry in the study of unpigmented planktonic protozoa is to a large extent unexplored. The goal of this study was to develop a protocol for single cell live sorting of freshwater ciliates which can be applied in future laboratory experiments. Unstained and DAPI-stained specimens of four species of common prostomatid freshwater ciliates were sorted and their survival and growth were then monitored in micro-well plates. Three out of four species investigated (Urotricha farcta, Urotricha castalia, and Coleps spetai) grew after sorting. For the two Urotricha species up to 92-100% of the sorted cells grew after sorting, depending on DAPI-concentration. For C. spetai up to 25% grew. The smallest and most delicate species, Balanion planctonicum, was difficult to sort unstained and appeared not to grow after single cell sorting. DAPI-staining at concentrations ranging from 0.1 to 5μg DAPIml-1 improved flow cytometric discrimination between the ciliates and non-target particles in the samples. The impact of DAPI-staining on the growth of the sorted cells differed between species. However, three of the four species tested grew reasonably well after staining with 0.1μg DAPIml-1. We expect that this staining and sorting protocol may greatly facilitate and improve future experiments with cultured ciliates.

Keywords

ciliatesflow cytometrylive sortingdna-staindapi.