Hi Everyone,

This will be the first time I will be fixing crustacean eggs for nuclear staining and as I will sampling at sea I would like to leave the eggs in the fixative solution for around 2 weeks before rinsing in buffer and staining. So my question is will leaving them in a fixative for a couple of weeks, rather than the 1h - 12h as I have seen in a handful of published articles, make a difference to the staining. The fixative is 97% glucamine-acetate buffer (GA buffer), 2% formalin, and 1% Triton-X (Sainte-Marie and Carriere, 1995). Where the GA buffer is composed of 250 mM N-methyl glucamine, 250 mM potassium gluconate, 50 mM Hepes, and 10 mM EGTA, adjusted to pH 7.4 with glacial acetic acid. https://spo.nmfs.noaa.gov/sites/default/files/pdf-content/1995/934/sainte-marie.pdf

Samples will be rinsed in a solution of phosphate buffered saline, 0.4% Triton X-100, 0.1% sodium azide and then placed in Accutase enzyme solution before staining.

Any insight would be greatly appreciated!

Many thanks!