I would like to see Gus expression in embryo sac and ovular tissue. So I need your suggestion and tricks for staining and clearing of tissue where expression of reporter gene desired. I usually do first gus staining then clearing by alcohol for seeing gus stain but by this way I didn't get stain anywhere even in desired tissue also. So I doubt on the penetrating limit of GUS stain solution in fresh tissue, so please suggest to me that, would it be better or not to fix and clear tissue first and then go for staining? The enzyme required gus staining will remain active or not after clearing before staining? If anybody has protocol for fixing and clearing of floral material of Arabidopsis please share with me.