Reference - Detail
|Author||Yamazaki T, Hirai C, Ota S, Kuwano K, Kuwano S.|
|Title||Use of FM1-43, a membrane-specific fluorescent dye, to estimate plasma membrane integrity in the cryopreservation of green algae.|
BACKGROUND AND OBJECTIVE:Cryopreservation of microorganism cultures is an important technology for their use as biological and genetic resources; however, the procedure is complicated and depends on the species.
MATERIALS AND METHODS:We used the two-step freezing method for the cryopreservation of the green alga Parachlorella kessleri.
RESULTS AND CONCLUSION:The optimal cryoprotectant for cryopreservation was 5% dimethyl sulfoxide plus 5% ethylene glycol. This is different from the optimal cryoprotectant for the closely related species Chlorella vulgaris. Efficient cryopreservation of P. kessleri was achieved using methanol, similar to Chlamydomonas reinhardtii. A membrane-specific fluorescent dye, FM1-43, was applied to estimate plasma membrane integrity. We found that the plasma membrane integrity of P. kessleri cells after freeze-thawing was associated with survivability, suggesting that this is a useful index for the optimization of the first step of the two-step freezing method of cryopreservation.
|MeSH||Cell Membrane / chemistry Chlorophyta / chemistry Chlorophyta / cytology* Cryopreservation* Cryoprotective Agents / chemistry Dimethyl Sulfoxide / chemistry Ethylene Glycol / chemistry Fluorescent Dyes / analysis* Freezing Pyridinium Compounds / analysis* Quaternary Ammonium Compounds / analysis*|
|WOS Category||BIOLOGY PHYSIOLOGY|