Publicaties | Kaarten

nieuwe zoekopdracht

[ meld een fout in dit record ]mandje (0): toevoegen | toon Print deze pagina
Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry
Randall, C.J.; Speaks, J.E.; Lager, C.; Hagedorn, M.; Llewellyn, L.; Pulak, R.; Thompson, J.; Bay, L.K.; Mead, D.; Heyward, A.J.; Negri, A.P. (2020). Rapid counting and spectral sorting of live coral larvae using large-particle flow cytometry. NPG Scientific Reports 10(1): 11 pp. https://dx.doi.org/10.1038/s41598-020-69491-0
In: Scientific Reports (Nature Publishing Group). Nature Publishing Group: London. ISSN 2045-2322; e-ISSN 2045-2322
Peer reviewed article  
Beschikbaar in  Auteurs 
Auteurs  Top 
  • Randall, C.J.
  • Speaks, J.E.
  • Lager, C.
  • Hagedorn, M.
  • Llewellyn, L.
  • Pulak, R.
  • Thompson, J.
  • Bay, L.K.
  • Mead, D.
  • Heyward, A.J.
  • Negri, A.P.
Abstract
    Research with coral embryos and larvae often requires laborious manual counting and sorting of individual specimens, usually via microscopy. Because many coral species spawn only once per year during a narrow temporal window, sample processing is a time-limiting step for research on the early life-history stages of corals. Flow cytometry, an automated technique for measuring and sorting particles, cells, and cell-clusters, is a potential solution to this bottleneck. Yet most flow cytometers do not accommodate live organisms of the size of most coral embryos (> 250 µm), and sample processing is often destructive. Here we tested the ability of a large-particle flow cytometer with a gentle pneumatic sorting mechanism to process and spectrally sort live and preserved Montipora capitata coral embryos and larvae. Average survival rates of mechanically-sorted larvae were over 90% and were comparable to those achieved by careful hand-sorting. Preserved eggs and embryos remained intact throughout the sorting process and were successfully sorted based on real-time size and fluorescence detection. In-line bright-field microscopy images were captured for each sample object as it passed through the flow-cell, enabling the identification of early-stage embryos (2-cell to morula stage). Samples were counted and sorted at an average rate of 4 s larva−1 and as high as 0.2 s larva−1 for high-density samples. Results presented here suggest that large-particle flow cytometry has the potential to significantly increase efficiency and accuracy of data collection and sample processing during time-limited coral spawning events, facilitating larger-scale and higher-replication studies with an expanded number of species.
Alle informatie in het Integrated Marine Information System (IMIS) valt onder het VLIZ Privacy beleid Top | Auteurs 
IMIS is ontwikkeld en wordt gehost door het VLIZ.