Anaerobic incubatorIt is a 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 in this device can cultivate the most difficult to grow anaerobic organisms, while avoiding the risk of anaerobic organisms dying from exposure to oxygen during previous operations in the atmosphere. Therefore, this device is an ideal tool for anaerobic biological detection research. This device is also a versatile instrument that provides favorable growth conditions for the growth and reproduction of microaerophilic bacteria by changing the operating mode.

The characteristics of anaerobic incubators are as follows:
1. Use scientific methods to achieve high precision in anaerobic environments, with good constant performance and reliable use;
2. The temperature of the incubator is controlled by a microcomputer, which can accurately and intuitively reflect the temperature inside the incubator, and can automatically control the temperature again. It is an effective temperature limiting protection device to ensure that the culture grows under safe temperature conditions;
3. The box is equipped with ultraviolet sterilization lamps, which can effectively prevent bacterial contamination;
4. The gas path device can accurately adjust the flow rate and input various required gases at will;
5. The pneumatic circuit is controlled by a microcomputer, and the solenoid valve can be started or closed with flexible operation;
6. The front window of the operating room is made of thick transparent special glass, which allows for clear and direct observation of indoor operations. The use of plastic gloves is reliable, comfortable, flexible, and easy to use;
7. There is a deoxygenation catalyst installed indoors.

Anaerobic incubatorMain technical parameters:
1. The anaerobic state in the sampling room takes less than 5 minutes to form;
2. The anaerobic formation time in the operating room is less than 1 hour (pumping and inflation replacement);
3. The anaerobic environment in the operating room should be maintained for more than 10 hours without replenishing trace amounts of mixed gas;
4. Operating temperature: Room temperature 3 ℃~50 ℃;
5. The temperature fluctuation in the cultivation room is ≤± 0.3 ℃; Temperature distribution uniformity ≤± 1.0 ℃;
6. Operating room volume: 820mm x 550mm x 660mm (length x height x width),
7. Incubator size: 250mm × 190mm × 290mm
8. Power supply 220V ± 10% AC, power: 500VA

Installation of anaerobic incubator:
1. The whole machine should be placed in a location with a small temperature difference and easy operation, avoiding direct sunlight and staying away from heating equipment, and placed steadily;
2. Place the mixed gas cylinder and nitrogen cylinder steadily, and install pressure reducing valves (including pressure gauges) in appropriate positions;
3. Connect the air circuit and check it. To prevent air leakage, if necessary, use sealant to bond at each connection point;

use

（1） Formation of anaerobic environment in the operating room:

1. Place necessary accessories and tools according to usage requirements;
2. Power on the lighting, turn on the temperature controller, and adjust the desired temperature;
3. Place 1000g of palladium particles (sealed) and 500g of desiccant in the operating room, and put in Meilan indicator paper (sealed);
4. Close the indoor and outdoor doors of the sampling room tightly and perform vacuum calibration;
5. Number of replacements in the operating room (nitrogen replacement):
A. Put latex gloves on the flange ring of the observation board and tighten them tightly
B. Connect the nitrogen intake path, open nitrogen control valve 1, make the gloves bulge, close valve 1, and then tighten the bag opening;
6. Repeat the nitrogen filling process for the second replacement of the operating room (nitrogen replacement), evacuate the sampling room first, and pay attention to using the foot switch to open and close the exhaust at any time. Three consecutive nitrogen replacements.
7. Fourth replacement of the operating room (mixed gas replacement):
(The mixed gas ratio is N2 85%, H2 10%, CO2 5%)
A. Switch the air circuit and open the mixing valve 3 for intake. When inflating, evacuate the sampling chamber first and use the foot switch to open and close the exhaust at any time;
B. Close the mixed gas valve 3 and open valve 5 to allow the mixed gas to pass through the flow meter. Adjust the flow meter to a flow rate of approximately 10ml per minute;
C. The mixed gas is repeatedly converted 2-3 times, and through the above process, an anaerobic environment is basically formed.
8. Turn on the granular 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 paper to observe its color change. If it does not change color, it means that the operating room has reached an anaerobic environment;
9. Turn on the ultraviolet sterilization lamp and sterilize indoors. The sterilization time should be determined according to the specific experiment.

（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;
3. Three rounds of nitrogen replacement process in the sampling chamber: Turn on the vacuum pump, first draw a vacuum degree of 500ml mercury column (66kPa) or above, stop, then manually open nitrogen valve 2 to intake air, make the pointer return to zero position, and close valve 2. Repeat the operation once for the second time. During the third operation, stop at a vacuum level of 500ml mercury (66kPa) or higher, then open valve 4 to introduce mixed gas and return the pointer to zero position, and close valve 4. The three nitrogen replacement processes in the sampling room have been completed;
If a lower vacuum degree is selected, the number of replacements needs to be increased;
5. Open and close the outdoor door for sampling, and then lower the vacuum level by 100ml of mercury (13kPa) for testing;
6. Conditions for long-term continuous use of anaerobic incubators:
A. Open the Meilan indicator paper in the operating room every day to observe. If it is not normal, it must be ventilated again;
B. Long term continuous input of trace amounts of mixed gas is required to enable the nitrogen gas to combine with trace amounts of oxygen through catalytic absorption, ensuring anaerobic conditions indoors. The flow rate of the mixed gas is selected to be around 10ml per minute;
C. Continuously cultivate and run for one day, and replace the deaerator and desiccant once.
D. The temperature inside the operating room can be freely selected and controlled.
7. Adjustment of output pressure for mixed gas cylinders and nitrogen cylinders: Adjust the pressure reducing valve to achieve an output pressure of around 0.1Mpa.

Notes:
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;