黄色小说视频-日本少妇高潮抽搐-国内自拍av-天堂中文资源在线-蜜桃视频在线入口www-91久久综合-亚洲视频在线一区-日日夜夜拍-国产精品资源在线观看-欧美激情国产精品免费-人妻少妇久久中文字幕-成人免费无码大片a毛片-香蕉污视频在线观看-日本电影成人-国内精品久久99人妻无码

Harvest and Preservation

1. Harvest cells from cultures which are at or near peak density.  Aseptically transfer cells to 15 mL plastic centrifuge tubes and centrifuge at ~150 x g for 5 min.
2. Adjust the concentration of cells to 2 x 10⁶/mL with fresh medium, then dilute to half this concentration by adding an equal amount of a 15% (v/v) sterile glycerol solution in fresh  medium.
3. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).  The time from mixing of the cell suspension and the glycerol solution, before the cooling cycle begins, should be no less than 15 min and no greater than 30 min.
4. Place vials in a controlled-rate freezing unit. From room temperature, cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)   
5. The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80 and -70°C for no longer than one week.
6. To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.
7. Immediately after thawing, do not leave in the water bath; aseptically remove the contents of the ampule and transfer to a fresh test tube (or T-25 flask) containing 5 mL of ATCC Medium 460. Incubate culture at 20-25°C (test tubes are incubated upright with the cap loosened one-half turn).  Subculture every 10-14d.

Japanese Patent 7,008,221

Salt GW. Changes in the cell volume of Didinium nasutum during population increase. J. Protozool. 22: 112-115, 1975.

Beam CA, Himes M. Sexual isolation and genetic diversification among some strains of Crypthecodinium cohnii-like dinoflagellates evidence of speciation. J. Protozool. 24: 532-539, 1977.

Seafood products having good taste containing marine microalgae, particularly Crythecodinium cohnii containing docosahexaenoic acid. Japanese Patent 7,008,221

Iizuka T, et al. Marine micro-algae food material containing docosahexaenoic acid, food containing the same and manufacturing method therefor. US Patent 5,547,699 dated Aug 20 1996

Beam CA, Himes M. Sexuality and meiosis in dinoflagellates. In: Beam CA, Himes M, Biochemistry and physiology of protozoa, 2nd ed.3 New York Academic Press: 171-206, 1980.