For visualisation, sections were incubated for 30 minutes in streptavidin-peroxidase complex (1 500; Dako), rinsed in distilled water, and incubated in fast-DAB (3,3'-diaminobenzidine) peroxidase substrate (Sigma-Aldrich) for 5 minutes and counterstained in diluted filtered haematoxylin (Sigma-Aldrich) for 15 seconds.
For visualisation, sections were incubated for 30 minutes at room temperature in streptavidin peroxidase complex (1 500 in DPBS; Dako), rinsed in distilled water and incubated in fast-DAB (3,3'-diaminobenzidine) peroxidase substrate (Sigma) for 5 minutes and counterstained in diluted filtered haematoxylin (Sigma) for 15 seconds.
Extra-avidin peroxidase (EAP) (Sigma, St . Louis MO, USA) with diaminobenzidine (DAB) (Vector Laboratories, Burlingame, CA, USA) as substrate were used for visualisation and sections were then counterstained with haematoxylin, washed, dried and mounted with buffered glycerol.
For visualisations, the sections were then incubated with 3-amino-9-ethylcarbazole (AEC) high-sensitivity substrate chromogen from DakoCytomation (Carpinteria, CA, USA) for 2 30 min.
During LMD due to the absence of cover slip over the tissue section visualisation of the morphology of the tissue becomes difficult therefore to facilitate identification of the morphology the tissue sections needs to be stained this can affect RNA viability.
Toluidine blue stained resin sections for visualisation of transverse vascular bundle sections in inflorescence stems were taken from 5 week old plants.
Following visualisation with diaminobenzidine, sections were counterstained with haematoxylin, dehydrated and mounted.
DNA quality was assessed by DNA extraction from a single tissue section and visualisation by agarose gel electrophoresis.
