Clearing Arabidopsis non-ovule materials with the HCG solution

Ref: Sabatini, S. et al., Cell 99, 463-472.

This protocol can be used for clearing intact non-ovule materials of arabidopsis, which can then be observed under Normarski optics. This is an efficient way to analyse root, seedling even flower development without sectioning.  This protocol could also be used for clearing GUS stained material, after chlorophyll is removed by 70% ethanol.

  1. To prepare the modified HCG solution:
    bullet30ml H2O 
    bullet80g Chloral hydrate (Sigma, C8383)
    bullet10 ml 100% Glycerol (Sigma, G6279)
    bulletDissolve by stirring for 1 hr.
  2. Put fresh materials to 2-4 drops of HCG solution.
  3. Gently cover the sample with a cover slip.
  4. Keep the slides at room temperature for 15' to overnight depending on the developmental stages of the material (younger materials clear quicker).
  5. Observe the samples with a microscope equipped with Nomarski optics.