Clearing arabidopsis seeds with Hoyer's
Clearing arabidopsis seeds with Hoyer's
This protocol can be used for clearing intact seeds of arabidopsis, which can then be
observed under Normarski optics. This is an efficient way to analyse embryo and endosperm
development without sectioning
- To prepare the modified Hoyer's solution:
- 7.5g gum arabic (Sigma, G9752)
- 100g chloral hydrate (Sigma, C8383, controlled substance)
- 5ml glycerol (Sigma, G6279)
- 60ml water
- dissolve by stirring for 3-5 hrs, or overnight.
- Open the siliques with a pair of forceps and a sharp needle, pick up the intact ovules with the needle.
- Put 2-3 drop of Hoyer's on a slide.
- Drop the ovules into the Hoyer's. 20-30 ovules for each sample.
- Gently cover the sample with a cover slip.
- Keep the slides at room temperature for 15' to overnight depending on the developmental stages of the ovules.
- Observe the samples with a microscope equiped with Nomarski optics.