The first step is to form the anaerobic environment of the cultivation operation room:
1. According to the requirements of use, place a deaerator and a closed culture dish in the cultivation operation room. 500g palladium particle oxygen scavenger (sealed state) and 500g desiccant.
2. Press the power switch, close the inside and outside doors of the sampling chamber, and press the vacuum pump button to draw the sampling chamber into negative pressure (see the pointer of the vacuum gauge)
3. To achieve a very small amount of oxygen in the cultivation operation room, three gas replacements must be performed: (1) the first gas replacement,
Nitrogen with a purity of 99.99%. The operation is as follows:
Put the latex gloves on the flange ring and tighten it. Press the nitrogen inlet switch in the box, the nitrogen will reach a sufficient state, and then press this switch to close it. Put your hands into the latex gloves, and then step on the foot switch. Nitrogen will gradually fill the culture chamber. (2) For the second gas replacement (nitrogen replacement), repeat the nitrogen filling process. The sampling chamber is evacuated first, and the exhaust switch must be opened and closed at any time. (3) The third replacement of the mixed gas, the mixed gas ratio is: N2 ↑ 90%, H2 ↑ 5%, CO2 ↑ 5%, and the purity is 99.99%. Vacuum the sampling chamber until the pointer of the vacuum gauge points to 0.06MPa. Press the mixed gas inlet switch in the box, the mixed gas reaches a sufficient state, and then press this switch to turn off. Press the mixed gas fine-tuning switch in the box to make the mixed gas fill the cultivation chamber at a certain flow rate. After three ventilations, the oxygen content of the gas in the cultivation operation room is already in a very trace state.
4. Turn on the palladium particle deaerator in the cultivation operation room, and turn on the power of the deaerator catalyst for catalytic deaeration.
5. If the user needs to operate the cultivation room or sampling room for sterilization before or after use, they can open the external door of the built-in incubator, that is, the sampling room door, and then turn on the control panel sterilization switch to sterilize the operation room. The length of time can be customized according to the actual needs of users. For details on the precautions for operating this function, please refer to Article (7) of the systematic “Precautions”.
The second step is to insert and cultivate the bacteria
1. Check the door of the sampling chamber and close it tightly.
2. Open the outdoor sampling door and close the external door after placing the bacteria in the sampling room.
3. Complete the nitrogen filling and replacement process of the sampling chamber three times: Turn on the vacuum pump, first evacuate to above 0.06MPa, and then manually turn on the aisle nitrogen inlet switch. When the pointer returns to zero, turn off the aisle nitrogen inlet switch. Repeat the above operation for the second nitrogen replacement. During the third gas replacement operation, stop when the vacuum degree is above 0.06MPa, and then turn on the aisle mixed gas inlet switch to charge the mixed gas. When the pointer returns to zero, turn off the aisle mixed gas inlet switch. The sampling chamber is aerated and replaced three times.
4. In order to ensure that the anaerobic environment of the cultivation operation room is not damaged, the replacement frequency should be increased when selecting a lower vacuum for nitrogen replacement.
5. Open the door of the sampling room, move the bacteria from the sampling room to the cultivation operation room, close the door of the sampling room tightly, and then vacuum check
Check if the inner door is closed tightly.
6. If the incubator needs to be used continuously for a long time, you must: (1) Observe in the cultivation operation room every day. If it is found to be abnormal, you must replace the gas. (2) When the user is operating, due to the product structure, after the gas supply is stopped for 12 hours, there may be a very small amount of leakage, or when the microbial gas supplement is required for the cultivation of the required bacteria, the built-in The filter continuously inputs a small amount of mixed gas, so that the supplemented nitrogen can be combined with a small amount of oxygen and absorbed by the catalyst to ensure the anaerobic state in the box. The supplementary mixed gas flow is about 10ml per minute. (3) Continuous culture for one day, replace the oxygen scavenger and desiccant once.
7. The temperature in the cultivation operation room can be arbitrarily selected and controlled within the temperature control range.
8. Adjusting the output pressure of the mixed gas bottle and nitrogen bottle: adjust the pressure reducing valve so that the output pressure is about 0.02Mpa.
Precautions and product maintenance and care:
1. The instrument should be installed in a place with clean air and small temperature change as much as possible.
2. Before starting, you should be fully familiar with and understand the instruction manuals of the supporting instruments and meters, and master the correct use methods.
3. The culture must be placed after the culture operating room has reached an absolutely anaerobic environment.
4. In the event of a failure (cause of gas outages, etc.), the culture operation room can still maintain an anaerobic state for 12 hours. (More than 12 hours, the culture needs to be taken out for further processing).
5. Always pay attention to the air leakage.
6. When the gas in the gas cylinder is exhausted and the total output pressure is less than 0.1Mpa, the gas cylinder should be replaced as soon as possible. When changing the gas cylinder, pay attention to tighten the gas pipe to prevent the oxygen gas from flowing into the pipe.
7. When the germicidal lamp needs to be turned on, the germicidal lamp tube can emit strong 2537Å short-wave ultraviolet rays, which has a particularly strong effect on nucleic acid proteins. Can make bacteria mutate and kill, at the same time, ultraviolet rays have lethal power to organic cells. Long-term exposure of human eyes and skin to ultraviolet rays can cause burns. Therefore, the culture in the operating room or culture room should be taken out when it must be turned on. After opening, you should leave the ultraviolet radiation in time to avoid discomfort, such as headache, chest tightness, dizziness and so on.
8. There is no power in the whole machine, you can check whether 220V power is plugged in at the power plug-in end. If there is power, unplug it and check whether the fuse on the control panel is blown. If the fuse needs to be replaced or repaired by a person with electrical or electrical knowledge, you can also consult the manufacturer. Do not replace it by yourself to avoid accidents and unnecessary losses.
9. When the initial set of gas is replaced or the mixed gas is released at the end of the culture, the doors and windows of the laboratory or the vents should be opened to accelerate the gas circulation. 。 Reduce the release of gas into the body and cause discomfort .