Precautions for anaerobic incubators

Anaerobic incubator is an experimental device for bacterial cultivation and operation under anaerobic environmental conditions. It can provide strict anaerobic conditions with constant temperature cultivation and has a systematic and scientific working area. Operating and cultivating anaerobic organisms in an anaerobic incubator can cultivate difficult to grow anaerobic organisms while avoiding the risk of anaerobic organisms dying due to exposure to oxygen during atmospheric operations. Therefore, this device is an ideal tool for anaerobic biological detection and research. This device is also a versatile and good instrument. If the operation mode is changed to provide good growth conditions for the growth and reproduction of microaerophilic bacteria, it can be equipped with built-in componentsOxygen detector (for self configuration), equipped with a flow control device on the operation panel for detection and necessary adjustment. Inputting the specified oxygen content for microaerophilic bacteria can provide favorable growth conditions for their growth and reproduction.

1. Install the instrument in a place with clean air and minimal temperature changes as much as possible;

2. Before starting up, it is necessary to fully familiarize and understand the instructions for using the various components and supporting instruments, and master the correct usage methods;

3. The culture must be placed in the operating room after reaching an anaerobic environment;

4. In case of malfunction (such as gas outage), the operating room can still maintain anaerobic state for 10 hours (if it exceeds 10 hours, the culture should be taken out for further treatment as needed);

5. Regularly pay attention to whether there is any air leakage in the air circuit;

When replacing gas cylinders, be careful to tighten the trachea to avoid the flow of oxygen-containing gases;

Use the vacuum pump as required and regularly check and refuel it.

1、 Formation of anaerobic environment in the operating room

Place the necessary accessories and tools according to the usage requirements, and put two non-toxic plastic bags into the operating room.

Turn on the lighting and temperature controller with a 2-channel power supply, adjust the required temperature and safe temperature.

Place 1000g of palladium particles (sealed) and 500g of desiccant in the operating room, and add Meilan indicator (sealed).

4. Close the indoor and outdoor doors of the sampling room tightly and perform vacuum calibration.

5. One replacement of DI in the operating room (nitrogen replacement):

（1) First, insert a rubber tube into the air inlet of the operating room, and then insert the other end into a plastic bag.

（2) Connect the nitrogen inlet, open the nitrogen control valve, ensure that the two plastic bags are filled with sufficient nitrogen, and then tighten the bag opening.

（3) Put latex gloves on the flange ring of the observation board and tighten them tightly.

（4) Gradually release the nitrogen gas inside the plastic bag into the operating room until it is completely released.

Repeat the nitrogen filling process for the DI secondary replacement (nitrogen replacement) in the operating room, and be sure to use the foot switch to open and close the exhaust at any time.

7 Operation room DI three times replacement (mixed gas replacement):

The mixed gas ratio is:N2↑ 85% H2 ↑ 10% CO2↑ 5%）

（1) Swap the air path and open the mixed air duct vent valve for intake. When inflating, always press the foot switch to open and close the exhaust.

（2) After filling the plastic bag with the mixed gas, turn off the mixed gas straight through valve (three-way valve), allow the mixed gas to pass through the flow meter input operation, and adjust the flow meter to a flow rate of about 10 milliliters per minute.

（3) Gradually discharge the mixed air inside the plastic bag into the operating room.

（4) After three rounds of ventilation, the oxygen content in the operating room gas has reached a very low level.

Open the palladium particle deoxygenation agent in the operating room, connect the power supply of the deoxygenation catalyst for catalytic deoxygenation, and after one hour, open the Meilan indicator (Meilan bottle) to observe its condition. If it does not change color, it indicates that the operating room has reached an anaerobic environment.

Turn on the UV sterilization lamp and sterilize indoors. Sterilization time can be determined by oneself.

2、 Placement and cultivation of bacterial strains

1. Check the door inside the sampling room and close it tightly.

2. Open the outdoor door of the sampling room, place the bacterial strain into the sampling room, and then close the door.

Three nitrogen replacement processes in the sampling chamber: first, evacuate to a vacuum degree of 500 mmHg (66Kpa) or above, then manually open the nitrogen valve to allow the pointer to return to zero before proceeding with the next operation.

If a lower vacuum degree is selected, the number of replacements needs to be increased.

5. Open and close the indoor and outdoor doors of the sampling room, and lower the vacuum level by 100 millimeters of mercury (13 Kpa) for inspection and assistance during operation.

6. Conditions for long-term continuous use of anaerobic incubators:

（1) Open the Meilan indicator in the operating room every day for observation, and use it normally. If it is not normal, it must be re ventilated.

（2) Long term continuous input of trace amounts of mixed gas is required to enable the added hydrogen to combine with trace amounts of oxygen through catalytic absorption, ensuring anaerobic conditions indoors. The flow rate of the added mixed gas is selected to be around 10 milliliters per minute.

（3) Continuously cultivate and run for one day, and replace the deaerator and desiccant once.

Application of 7 Inoculation Rod Sterilizer:

（1) Use nickel chromium electric heating wire at any time to short-circuit the electrode and make it red hot for sterilization (inoculation rod)

（2) The wax melting point can be directly placed on it, and the wax sealing mouth of the test tube can be rotated to melt the wax.

The temperature inside the incubator in the operating room can be freely selected and controlled.

Adjustment of output pressure for 9 mixed gas cylinders and nitrogen cylinders: Adjust the pressure reducing valve to achieve an output pressure of around 0.1Mpa.