Vision lite 2

Heart perfusion was conducted to excise the whole brain according to the procedure described by Gage et al. (2012) . Tissue sections (5 µm) of the hippocampus were deparaffinized with xylene, rehydrated, and washed with 70% isopropanol and double-distilled water (ddH₂O) respectively. The Congo red stain (Cat #C6767, Merck Germany; working solution: 49.5 ml Congo red (Stock) and 0.5 ml 1% NaOH) was poured on the deparaffinized brain sections and retained for 20 min. ddH₂O and alkaline alcohol were used to wash sections for 2 min. The sections were then counterstained by hematoxylin for 30 s and further washed with 70% isopropanol for 6 min and then with ddH₂O. After air-drying (1 h), the slides were mounted by coverslips and later visualized using B-150, OPTIKA microscope (Italy) at 40×, 10×, and 4× resolution. The images were captured using Optika Vision Lite 2.1 image analysis software.

Leaf disk of approximately 2–3 cm of both inoculated and non-inoculated banana leaves were detached and soaked in 10 ml of absolute HCl for 20 min. The samples were then washed in sterile distilled water for 5 min and soaked twice in 100% ethanol for 1 h to remove chlorophyll. Finally, the leaf samples were rinsed in sterile distilled water for 5 min, dehydrated with 95% ethanol and stored at 4°C until ready for staining.
To observe fungal mycelium growth and development within banana leaf cells, the clear leaves were stained with LPCB for 30 min. Excess stains on leaf disk were blotted and fixed on slides. The slides were observed under COSLAB light microscope and picture was taken using digital camera MDCE- 5C (ISO 9001 Co) and analyzed using Optika Vision Lite 2.1 software.