Equipment preparation: Check if the power supply and gas source (CO ₂ steel cylinder) connection of the CO ₂ shaker are normal, and ensure that the pressure of the CO ₂ pressure reducing valve is stable (usually 0.1~0.2MPa). Turn on the device power, preheat to the set temperature (such as 37 ℃), and preheat the water tray in the incubator to maintain humidity.

Parameter setting: Set the CO ₂ concentration (usually 5%) according to the cell requirements, and calibrate it through infrared or thermal conductivity sensors; The temperature range is generally set from room temperature to 50 ℃, and preheating is required in advance to avoid temperature fluctuations; Adjust the rotation speed (50-300rpm) and amplitude (usually 5-50mm) according to the culture container to avoid liquid splashing out; Add sterile distilled water to the water tray and maintain a humidity of ≥ 95%.

Sample placement: The container needs to be sealed (such as using breathable cap film or tightening the bottle cap) to prevent CO ₂ from escaping and contaminating; Balance the placement of samples to avoid center of gravity deviation affecting the operation of the shaker.

Operation monitoring: Regularly check the stability of CO ₂ concentration and temperature, and record operating parameters; When using breathable cap culture bottles or sealed bottles, it is necessary to balance the CO ₂ permeability.

End operation: Turn off the CO ₂ gas source and wait for the concentration to drop to 0% before shutting down to avoid sensor damage; Clean the water tray and chamber to prevent microbial growth.

2、 Best Practice Guidelines for Cell Culture

Cell type and culture medium selection: Choose the appropriate culture medium based on the cell type, such as DMEM for rapidly proliferating cell lines and MEM for multiple cell types; Primary cells require the addition of special growth factors.

Cell recovery and cryopreservation: Quickly thaw cells during recovery to reduce ice crystal damage; Add cryoprotectant (such as 10% DMSO) during cryopreservation, slowly cool to -80 ℃, and then transfer to liquid nitrogen.

Aseptic operation: All operations are carried out in an ultra clean workbench, wearing laboratory clothes, gloves, masks, and hats; Avoid speaking and coughing in the operating area to reduce the risk of microbial transmission through the air.

Regular observation and recording: Regularly observe cell growth under a microscope, including cell morphology, growth density, and contamination; Record relevant data for timely detection and handling of abnormal situations.

Equipment maintenance and calibration: Regularly calibrate the CO ₂ sensor (once every 6 months), replace the filter membrane (if any); Power off during cleaning to prevent liquids from entering the electrical system.