I need to know how to store Arabidopsis roots for further analysis in a way that hair roots are preserved in their integrity, without degrading or collapsing, suffering from dehydration, etc.
I will have too many hydroponic samples to analyse in a single day.
Therefore, I must store the samples to analyse them later on.
Only morphology interests me, not celular contents.
It would be very useful to have a reference, focused in Arabidopsis hair roots.
Many thanks in advance!
What are you hoping to study? The form of the hairs themselves? It's difficult to preserve both morphology and cellular contents (RNA etc). If it is morphology, my vote would be to do all your imaging with fresh live samples, then you can perform any image analysis later.
In my experience, root hairs just don't keep very well. They do image well when plants are grown vertically on MS plates with phytagel.
Thanks for your response!
I will have too many hydroponic samples to analyse in a single day.
Therefore, I must store the samples to analyse them later on.
Only morphology interests me, not celular contents.
Well, you could scale down your experiment or only do fine-scale imaging on a random subset of the answer plants and/or roots
or break up the experiment into subsets
I also think, that it is better to study morphology from fresh samples. If they are still sterile, You can store them dirrectly in Petri dishes with MS, or in sterile colba or bottles with liquid 1/2 MS in room temperature up to 2-3 weeks without transplantation. If they are not sterile in the moment, You can make solution of water with Cefotaxim, 500 mg/L wash them in this solution and put then roots into sterile liquit 1/2 Ms with 400 mg/l Cefotaxim. Store at 16-19 0C during several days. If You can do that analysis during two days, You can just store nonsterile hairy roots in water in dark not hot conditions.
Second link from my colleage, who during many years work with fixation of plant tissues, I have never tried his fixation method. In any way, You can take first small amount of roots and try before usage of this method for all summ of hairy roots.
https://pubmed.ncbi.nlm.nih.gov/29235975/
https://academic.oup.com/plcell/article/20/6/1494/6092150?login=true
Roots of Arabidopsis for morphological analysis can be spread on agar plates using a brush and can be stored at 4 degree for few days.
maybe, about that temperature 4 degree i don't know. What I wrote about storage at room temperature without fixation - I used previously fresh hairy roots of Amaranth for UV microscoping. I think better in liquig 1/2 Ms with antibiotic or water is better, they will be less distructed. for separating roots better to use needlke or scalpel. With brash, You can broken them.
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