| 产品名称： |
Willaertia magna De Jonckheere et al. |
| 商品货号： |
TS187677 |
| Deposited As： |
Protonaegleria westphali Michel |
| Strain Designations： |
NI4C1(1) |
| 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. |
| Product Format： |
frozen |
| Storage Conditions： |
Frozen: -70°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage |
| Axenic/Xenic： |
Xenic |
| Type Strain： |
no |
| Comments： |
ultrastructure |
| Medium： |
ATCC® Medium 997: Fresh water ameba medium
ATCC® Medium 711: PYB
|
| Growth Conditions： |
Temperature: 25°C |
| Cryopreservation： |
Cryoprotective Solution
DMSO, 1.5 ml
Pages Balanced Salt Solution (or similar), 8.5 ml
Harvest and Preservation
- Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.
- Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 1323 (Pages Balanced Salt Solution) and washing cells into suspension.xa0 Rub the surface of the plate with a spread bar to detach adhering trophozoites.
- Adjust the concentration of cells to at least 2 x 106/ml in fresh medium.
- Mix the cell preparation and the cryoprotective solution in equal portions.
- Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
- 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
- Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.
- To establish a culture from the frozen state place the vial in a 35°C water bath.xa0 Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.xa0xa0 Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.xa0 Distribute the material evenly over the plate using a spread bar.
- Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.
- Follow the protocol for maintenance of culture.
|
| Name of Depositor： |
R Michel |
| References： |
Michel R, et al. Ultrastructure of the amoebo-flagellate Protonaegleria westphali. Parasitol. Res. 74: 23-29, 1987. PubMed: 3438290
|
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