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The salinity of 53 was chosen to check for viability of planulae at the maximum salinity that the lagoon had before the connection to the Mediterranean Sea was enlarged.
To check for viability, cells were diluted 1 2 with trypan blue (LifeTechnologies).
Cell counts were measured using serial dilution colony counting before increasing hydrolysate concentration to check for viability.
To check for viability, slides were kept after imaging to confirm that embryos subsequently hatched as larvae.
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Cells were counted, checked for viability and resuspended in R10 at concentration of 10 6cells/ml.
In parallel, acridine orange stained cells were counted and checked for viability using fluorescent microscopy.
Cells were counted, checked for viability, and re-suspended in R10 at 106 cells/mL.
Both assays were incubated at 20°C and were checked for viability daily (for the paraquat test) or every hour (for the juglone test) by using a Nikon SMZ800.
To generate survival curves, eclosed flies were kept individually in small vials with standard food and checked for viability every morning.
Sporangia were checked for viability by germination assays [ 58].
Cells were checked for viability under the optical microscope.
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