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微信小章| 产品名称： | Tokophrya infusionum (Stein) Collin |
|---|---|
| 商品货号： | TS187050 |
| Strain Designations： | clone 25 |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Isolation： | pool at Rockefeller University, New York City, 1959 |
| Product Format： | test tube |
| Type Strain： | no |
| Comments： | Localization of acid phosphatase use of plastic ampoules for freeze preservation |
| Medium： | ATCC® Medium 875: AUY |
| Growth Conditions： | Max Temperature: 25.0°C Min Temperature: 19.0°C Duration: grown with Tetrahymena borealis ATCC 30321 Protocol: ATCCNO: 30297 SPEC: Tokophrya infusionum must be periodically fed Tetrahymena (e.g., T. borealis ATCC 30321). Two methods may be used: 1) Prior to subculturing, feed 0.03 ml of a 5- to 7-day-old culture of ATCC 30321 to T. infusionum. A large number of embryos will be formed 1-4 hours after feeding. Transfer 1 ml of medium containing the free-swimming embryos to new medium. The number of embryos is variable depending upon the state of the culture and the number of tetrahymenas fed. 2) If difficulty is encountered in using method 1, the following method can be used with satisfactory results: Place the stock culture in an ice bath for 15-20 min., then agitate it. Some adults will detach from the glass surface. Transfer 1.0-2.0 ml of the medium containing dislodged adults to fresh medium and immediately feed with 0.03 ml of Tetrahymena. Four to five days after subculturing, feed the cultures again with 0.3 ml of Tetrahymena. Cultures are usually maintained in 20 X 125-mm screw-capped tubes containing 15.0 ml, kept upright with caps loosened. Cells attach in the region of the meniscus. |
| Subcultivation： | Protocol: ATCCNO: 30297 SPEC: Tokophrya infusionum must be periodically fed Tetrahymena (e.g., T. borealis ATCC 30321). Two methods may be used: 1) Prior to subculturing, feed 0.03 ml of a 5- to 7-day-old culture of ATCC 30321 to T. infusionum. A large number of embryos will be formed 1-4 hours after feeding. Transfer 1 ml of medium containing the free-swimming embryos to new medium. The number of embryos is variable depending upon the state of the culture and the number of tetrahymenas fed. 2) If difficulty is encountered in using method 1, the following method can be used with satisfactory results: Place the stock culture in an ice bath for 15-20 min., then agitate it. Some adults will detach from the glass surface. Transfer 1.0-2.0 ml of the medium containing dislodged adults to fresh medium and immediately feed with 0.03 ml of Tetrahymena. Four to five days after subculturing, feed the cultures again with 0.3 ml of Tetrahymena. Cultures are usually maintained in 20 X 125-mm screw-capped tubes containing 15.0 ml, kept upright with caps loosened. Cells attach in the region of the meniscus. |
| Cryopreservation： | Cryoprotective Solution DMSO xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 2.0 ml Fresh growth medium w/o bacteriaxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 8.0 ml 1.xa0xa0xa0xa0 Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat. 2. xa0xa0 Harvest Tokophrya cells from a culture that has recently passed peak density by centrifugation at 250-300 x g for 5 min. 3.xa0xa0xa0xa0 Adjust the concentration of cells to at least 2 x 104/ml in fresh medium. 4.xa0 xa0xa0 Mix the cell preparation and the cryoprotective solution in equal portions by adding the cryoprotective solution to the cell suspension in 3 equal aliquots at 2 min. intervals. 5.xa0 xa0xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6. xa0xa0xa0 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.xa0 Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately -1°C/min.) xa0 7.xa0 xa0xa0 Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator. 8.xa0 xa0xa0 To establish a culture from the frozen state place the vial in a 35°C water bath. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial. Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and transfer to a petri plate or T-25 tissue culture flask containing a bed of non-nutrient agar (ATCC medium 919) and 10 ml ATCC medium 875 diluted in ATCC medium 1323. 9.xa0xa0xa0xa0 Aseptically transfer 0.2-0.5 ml of washed Tetrahymena to the petri plate or T-25 flask (see section on MAINTENANCE OF CULTURE).xa0 Incubate the culture at 20-25°C. Once the culture is established, follow the protocol for maintenance of culture. |
| Name of Depositor： | MA Rudzinska |
| Year of Origin： | 1959 |
| References： | Rudzinska MA. Ultrastructural localization of acid phosphatase in starved Tokophrya infusionum. J. Protozool. 21: 721-728, 1974. PubMed: 4217373 Simione FP Jr., et al. The use of plastic ampoules for freeze preservation of microorganisms. Cryobiology 14: 500-502, 1977. PubMed: 891238 |