Freeze-drying technique preserves the stability of nanoparticles. The objective of this study was optimization of freeze-drying condition of nano lipid carriers (NLCs). NLCs were prepared by emulsion-solvent evaporation followed by ultra-sonication method. Different carbohydrate and polymeric cryoprotectants including Microcelac® (mixture of lactose and Avicel), Avicel PH102 (microcrystalline cellulose), mannitol, sucrose, Avicel RC591 (mixture of microcrystalline cellulose and sodium carboxymethyl cellulose), maltodextrine, Aerosil and PEG4000 were tested initially. The NLCs showing lower particle size growth and greater absolute zeta potential after freeze drying were chosen for further investigation using Taguchi optimization method. Studied factors included cryoprotectant type and concentration, freezing temperatures applied at different time periods and sublimation time. Sucrose, Avicel RC591 and Aerosil were selected as cryoprotectants from initial screening tests. Increasing their concentration increased the particle size. 1% of Avicel RC591, 24 h of freezing at −70 °C and 48 h sublimation time showed lower growth in particle size.