First, the operation room anaerobic environment formation 1. Place the necessary accessories and utensils according to the use requirements, and put two non-toxic plastic bags into the operation room.
2. Turn on the light through the power supply, turn on the temperature controller, adjust the required temperature and safe temperature.
3. Place 1000 g of palladium pellets (blocked) and 500 g of desiccant in the operating chamber and place the Meilan indicator (closed).
4. Close the indoor and outdoor doors and vacuum check.
5. Operating room * replacement (nitrogen replacement):
(1) Insert the rubber inlet into the air inlet of the operating chamber and insert the other end into the plastic bag.
(2) Turn on the nitrogen inlet, open the nitrogen control valve, make the two plastic bags full of nitrogen, and then tighten the bag.
(3) Put the latex gloves on the flange of the observation plate and tighten them.
(4) The nitrogen in the plastic bag is gradually placed in the operating room until it is completely discharged.
6. The second replacement of the operating room (nitrogen replacement) repeats the nitrogen filling process, and pay attention to the opening and closing of the exhaust gas with the foot switch at any time.
7. The third replacement of the operating room (mixed gas replacement):
(The mixture ratio is: N2: H2: CO2 = 85%: 10%: 5%)
(1) Change the air passage to open the mixed air passage through the valve intake air. When inflating, the foot switch should be opened and closed at any time.
(2) After the mixture is filled with the plastic bag, turn off the mixture gas straight-through valve (three-way valve), make the mixture flow through the flow meter input operation and adjust the flow meter, the flow rate is about 10 ml per minute.
(3) The mixed air in the plastic bag is gradually discharged into the operating room.
(4) After three times of gas exchange, the oxygen content of the gas in the operating chamber is already in a very small amount.
8. Open the palladium particle deoxidizer in the operation room, turn on the deoxidation catalyst power supply to carry out catalytic deaeration, open the Meilan indicator (Merlan bottle) after one hour to observe the discoloration, and do not change color to achieve anaerobic operation in the operation room. surroundings.
9. Open the UV sterilization lamp and sterilize it indoors. The sterilization time is self-determined.
Second, the placement and cultivation of strains 1. Check the sampling room door and close.
2. Open the sampling outdoor door and close the outer door after placing the strain into the sampling room.
3. The sampling chamber is filled with nitrogen for three times: firstly, the vacuum is 500 mmHg (66Kpa) or more, and then the nitrogen valve is manually opened to make the pointer return to the zero position for the next operation.
4. If the selected vacuum is low, the number of replacements needs to be increased.
5. Sampling indoor and outdoor doors to open, close to vacuum and 100 mm Hg (13Kpa) inspection and help operation.
6. Anaerobic incubators require long-term continuous use conditions:
(1) Open the Meilan indicator in the operating room every day and use it under normal conditions. If it is not normal, you must re-ventilate.
(2) It is necessary to continuously input a small amount of mixed gas for a long period of time, so that the supplemental hydrogen gas can be combined with a trace amount of oxygen to pass through the catalytic absorption, thereby ensuring the indoor anaerobic state, and the flow rate of the mixed gas mixture is selected to be about 10 ml per minute.
(3) Continuously run for one day, and replace the oxygen scavenger and desiccant once.
7. Application of inoculation rod sterilizer:
(1) Short-circuit the electrode with a nickel-chromium heating wire at any time to become red hot for sterilization (inoculation rod)
(2) The molten wax can be placed directly, and the test tube is sealed with a wax to rotate the molten wax.
8. The temperature in the incubator in the operating room can be arbitrarily selected and controlled.
9. Mixing gas cylinder and nitrogen bottle output pressure adjustment: adjust the pressure reducing valve so that the output pressure is about 0.1Mpa
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