ISSN:
1550-7408
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Biology
Notes:
Abstract. A flow cytometry (FCM) assay using SYBRgreen and propidium iodide double staining was tested to assess viability and morphological parameters of Perkinsus marinus under different cold- and heat-shock treatments and at different growth phases. P. marinus meront cells, cultivated at 28°C, were incubated in triplicate for 30 min at −80°C, −20°C, 5°C, and 20°C for cold-shock treatments and at 32°C, 36°C, 40°C, 44°C, 48°C, 52°C, and 60°C for heat-shock treatments. A slight and significant decrease in percentage of viable cells (PVC), from 93.6% to 92.7%, was observed at −20°C and the lowest PVC was obtained at −80°C (54.0%). After 30 min of heat shocks at 40°C and 44°C, PVC decreased slightly but significantly compared to cells maintained at 28°C. When cells were heat shocked at 48°C, 52°C, and 60°C heavy mortality occurred and PVC decreased to 33.8%, 8.0%, and 3.4%, respectively. No change in cell complexity and size was noted until cells were heat shocked at≥44°C. High cell mortality was detected at stationary phase of P. marinus cell culture. Cell viability dropped below 40% in 28-day-old cultures and ranged 11–25% in 38 to 47-day-old cultures. Results suggest that FCM could be a useful tool for determining viability of cultured P. marinus cells.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1550-7408.2005.00058.x
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