Team:Jiangnan/Hardware
Device-plasma
Configuration
The plasma equipment consists of a helium bottle, a rotor flowmeter (which monitors gas flow), high and low voltage outlets, a power supply and an oscilloscope (which displays voltage and current conditions).
When using this device, we place samples under the ejection nozzle, open the helium bottle, turn the power until the rotor flowmeter reaches the desired parameters (0.6-0.8 m/h, 20℃, 101325 MPa) and the oscilloscope displays 110V.
Protocol
1 ml blank DMEM medium was added to the 12-hole plate and placed directly under the low temperature atmospheric pressure plasma generator. The distance between the end of plasma beam and the medium is 1 cm, the injection port is vertically aligned to the center of the hole in the 12-hole plate, the controlled helium flow rate is 1 slm / min, and the output voltage of the high frequency power supply controlled by the transformer is 0.96-1.24 Kv. Plasma activated medium (PAM) was obtained by repeating the aforementioned process. When cells were incubated with viruses, the active oxygen and the active nitrogen in the culture medium had a beneficial effect on virus replication. 500000 MDBK cells were placed in the 6-hole plate for 24 hours. Then the original culture medium was changed to 1 ml PAM (DMEM activated by plasma). IBRV were then added and incubated with MDBK cells for 1 hour before changing the medium to the maintaining 1640 medium (supplemented with serum at 2% final concentration). The culture was collected for virus titer detection after cell lysis.
Result
After incubating IRF7-silenced cells with the DMEM medium for 1 hour, IBRV replication in MDBK was further increased 2.5 folds.
Advantages of using plasma activated medium in cell treatment:
(1) The intensity is controllable and the results are stable.
(2) The approach is simple and fast, which takes about 2 min for the treatment.
(3) The efficacy is obvious, which easily increase virus titer 2.5 folds further.
Figure 1. Configuration of cold atmospheric plasma source.
Figure 2. Protocol of cold atmospheric plasma activated medium preparation.