Introduction of a temperature calibration method for living cell inverted fluorescence microscope

Live cell culture fluorescence microscope can be used for routine culture conditions of living cells, such as temperature control, humidity control, CO2 concentration control, O2 concentration control, and high-speed camera can be configured for time series (Time-Lapse), combined with multi-color fluorescent filter Switching, LED single-wavelength source excitation, can be equipped with phase difference (Phase Contrast), differential interference (DIC), fluorescence and other contrast methods can be more intuitive and detailed cell morphology record and is recognized by the majority of scientific research users. In recent years, experiments on the placement of live cell culture devices with inverted fluorescence microscopy have become more common. There are many integrated fluorescence imaging systems available on the market, such as GE's DELTAVISION live cell imaging system, ZEISS's CELL OBSERVER system, or other large The brand's complete live cell imaging system is expensive (over $200,000) and has high maintenance costs. Manufacturers' engineers often have slower speeds or expensive fees, which is difficult for routine laboratories. At present, each laboratory is usually equipped with an inverted fluorescence microscope. The living cell culture device is a modular design product. Guangzhou Keshi Special Science Instrument Co., Ltd. provides a convenient and simple solution to customize a complete set of living cells or existing fluorescence. Microscope for live cell upgrade, low cost of purchase, easy disassembly and assembly, compatible with the major microscope platforms currently in the market, such as: Leica, Nikon, Olympus, Zeiss, etc. can provide customized adapters.

Temperature control in live cell culture devices is an important experimental parameter, but because live cell culture detectors are usually located at the upper wall edge of the culture chamber, due to room temperature (eg 22 ° C) and set temperature in the incubator (eg 37 ° C) The deviation is relatively large, the temperature conduction heat dissipation effect, the true temperature of the cells in the microscope cell culture incubator is difficult to reach the set temperature, in order to ensure the true culture temperature is accurate, the microscope live cell culture usually needs to be carried out before the first use in different laboratory environments. Calibration.

Recently, the customer who installed the Guangzhou Keshi Special Science Instrument Co., Ltd. asked us about the temperature calibration problem. By the way, the IBIDI live cell culture device represented by our company is used as an example to introduce the temperature calibration of the live cell culture device. Guangzhou Keshi Special Science Instrument Co., Ltd. is the domestic authorized agent of IBIDI live cell culture products in Germany, with favorable price and fast delivery speed.

one. The composition of the microscope live cell device usually includes a living cell culture chamber, a control module unit, a humidification column, a connection catheter, a CO2 gas cylinder, a compressed air bottle and the like.

Second, the incubator cavity is covered by the upper cover (LID) and heated

The base plate (PLATE) is fixed on the stage of the inverted microscope by an adapter, and has a temperature probe, a gas input and output interface, and a control panel can set a gas flow rate, a CO2 and an O2 concentration.

three. Take out the calibration-specific gadget and calibrate the temperature probe, a cable that can be inserted into the inside of the culture dish. A calibration petri dish with an open top. Insert the probe into the small hole of the calibration dish and use this probe to monitor the temperature inside the culture dish. The goal is to set the internal temperature to the target cell temperature (eg 37 ° C).

four. Turn on the temperature controller power switch and display the relevant parameters on the front panel LED panel of the controller. The controller can control the temperature setting of up to 4 channels. The upper cover and bottom plate of the live cell culture device can be heated. Usually only the first two channels need to be occupied. The temperature of the internal probe indicated by I, the set temperature indicated by S, E Indicates the temperature of the external probe. For example, if we want the temperature in the living cell culture dish to be 37 ° C, we can adjust the set temperature of the upper cover (channel 1) and the low heating plate (channel 2), when the external probe temperature indicated by E reaches 37 ° C, and It lasts for 15 minutes and remains unchanged. The calibration is completed.

Fives. After the calibration is completed, record the set temperature of the upper cover (LID) and the lower heating plate (PLATE). Each time you start the experiment, you only need to keep the set temperature at the temperature we have already calibrated. It can be considered that the temperature of the experimental environment inside the dish can be kept at the target temperature, for example, 37 °C. This process is similar to the pre-experiment performed prior to performing live cell culture experiments.

six. Inverted fluorescence microscope live cell culture modular design, this device can be compatible with the major microscope brands, if the laboratory has more than one brand of inverted microscope, only one can be used with the installation, convenient and practical. If you have any questions, you can consult the service engineer of Guangzhou Keshi Special Scientific Instrument Co., Ltd.