Hello,
I am optimizing a protocol to visualize pollen tube growth in several grass species. I am using ethanol-glacial acetic acid to fix the flowers and then plan to re-hydrate them using serial ethanol solutions (70%, 50%, 30%, water). Then I will clear the tissue using a 8M Sodium Hydroxide solution. I was wondering which steps are time-sensitive and which ones are not. I think I have to keep the flowers in ethanol-glacial acetic acid for a certain amount of time (I am trying several concentration-duration combinations). I believe I can leave the samples in 70% ethanol for weeks without problem, but not in the Sodium Hydroxide solution. So after the clearing, if I want to storage the flowers for some time, what solution should I use? I am using aniline blue staining, would it be good to keep it in the staining solution for ~1-2 weeks?
Thank you in advance for your help.
Dear Lara, sorry to see that your very interesting technical question has not yet received any expert answers. As an inorganic chemist I am unfortunately not an expert in this field. However, I suggest that you search RG for possible answers, particularly the "Publications" and "Questions" sections. For example, I just came across the following closely related question which has been asked earlier on this platform:
Why is 70% ethanol used to store samples for genetic studies?
https://www.researchgate.net/post/Why_is_70_ethanol_used_to_store_samples_for_genetic_studies
(19 answers)
Chances are that you will find potentially useful answers to your question right here on RG. Good luck with your work!
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