The cell three gas incubator provides an in vitro culture environment close to physiological conditions for adherent cells, stem cells, etc. by precisely regulating the concentrations of CO ₂, O ₂, and N ₂. The correct use of it is directly related to the reliability of experimental data and the success rate of cell culture. The following provides a systematic explanation from the operation process to maintenance:

1、 Preparation before use

1. Environmental verification

Confirm that the device is placed on a horizontal table with a reserved heat dissipation space of ≥ 30cm around it, away from heat sources and direct sunlight;

Check if the power supply voltage matches the value indicated on the equipment nameplate, and ensure that the grounding wire is reliably connected;

Conduct a no-load operation test before activation to verify the stability of the temperature control system (set at 37 ℃) and CO ₂ control system.

2. Cabin pretreatment

After new equipment or long-term disuse, use 75% ethanol to wipe the inner walls and tray supports, and sterilize them with UV for 30 minutes;

Pre balanced gas environment: After closing the box door, introduce a mixed gas (such as 95% air+5% CO ₂) and run for 2 hours to stabilize the internal pressure.

2、 Core parameter settings

1. Temperature and humidity management

Temperature setting: Most mammalian cells are suitable for 37 ± 0.5 ℃, and the actual temperature is recorded every hour after startup;

Humidity control: Maintain a saturation humidity of 95% ± 3%, add sterile water to the wet tray daily to avoid condensation droplets falling.

3、 Operating standards

1. Sample insertion and extraction

Rapid opening and closing of doors can cause a sudden change in pressure inside the box, and access should be completed within 1 minute;

The culture bottles/dishes should be evenly distributed on the tray to avoid blocking the circulation air outlet;

The operator wears sterile gloves and the tools are sterilized by an alcohol lamp flame.

2. Daily monitoring

Check the real-time parameters on the display screen daily, with a focus on observing:

-Consistency between CO ₂ sensor reading and infrared detection window;

-Does the temperature fluctuation curve exceed ± 0.3 ℃;

-Fill the water level line of the water tank with sterilized injection water in a timely manner.

4、 Maintenance and upkeep

1. Periodic maintenance

Weekly: Clean the dirt at the gap of the door seal and test the alarm function (cut off the power to simulate a fault);

Monthly: Calibrate the CO ₂ infrared sensor and replace the HEPA filter at the air inlet;

Quarterly: Drain the humidifier water tank and disinfect the entire cabin with 1% sodium hypochlorite solution.

2. Emergency response

Immediately cut off the main power supply and open the pressure relief valve to reduce the pressure in the box when the overheating alarm occurs;

Stop the experiment when pollution occurs, remove all items and perform formaldehyde fumigation disinfection.

5、 Key precautions

Frequent opening and closing of the box door is prohibited, and it is recommended to observe it no more than 3 times a day;

Do not stack multiple layers of shaking tables or heavy equipment to avoid affecting airflow distribution;

When not in use for a long time, drain the water tank, set the humidity to 0%, and run it for 1 hour every month to prevent moisture.

The refined management of the three gas cell incubator requires a balance between equipment characteristics and biological requirements. Through standardized operating procedures, strict parameter monitoring, and regular maintenance, the quality of cell culture can be significantly improved, providing a stable experimental platform for drug screening, tissue engineering, and other research.