Abstract
Cryopreservation of sporothalli of the red alga Porphyra (P. seriata, P. yezoensis, P. tenera, P. pseudolinearis and P. dentata) was performed by the two-step cooling method in liquid nitrogen. The algal samples were suspended in various cryoprotective solutions, and slowly cooled to -40 in 4 hours using a programmed freezer. After the first slow cooling the suspensions with cryoprotectants were immediately immersed in liquid nitrogen. The suspension from the programmed freezer was thawed quickly by agitation of the vial in a water bath at 40°C. When ice in the suspension of cryogenic vial was mostly melted, the vial was transferred to an ice bath for complete melting of the residual ice. The cryoprotectants in the vial were washed off by gradual dilution with seawater. The viability of the cell was assessed with neutral red staining. The viability of Porphyra samples ranged 54.6-70.9% when the mixed suspension of 10% dimethylsulfoxide and 0.5 M sorbitol in 50% seawater used as a cryoprotectant.
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