Our aim was to entrap Amphotericin-B, within the nanoparticles so that it could penetrate the cornea effectively. Solvent displacement method was used to prepare the nanoparticles. Confocal laser scanning microscopy was used to check the corneal penetration of the nanoparticle into isolated goat eyes after contact time of 1 hr and 24 hrs with the formulations. Prepared nanoparticles had particle size range from 112 to 350 nm with a zeta potential of 19 to 34 mV. This positive zeta helps the nanoparticle to adhere with corneal mucosa and release the drug. Almost 60-70% of the drug was released from every formulation in 6 to 7 hrs. The nanoparticles were able to entrap upto 75% of the drug successfully. Ex-vivo study reveals that 50% of the topically applied drug penetrated the cornea within 24 hrs whereas the plain drug solution had negligible penetration capacity.