The cell environment imaging system can monitor the growth status, morphological changes, and metabolic activity of cells in real time, helping researchers to more accurately understand the physiological state and response of cells. By regularly recording and analyzing image data, abnormal situations such as cytotoxicity, malnutrition, or pathogen infection can be detected in a timely manner, and corresponding measures can be taken to optimize cell culture conditions or solve problems.
The basic working principle of cell environment imaging system:
1. Optical imaging technology: Cell environment imaging systems often use inverted microscope structures, integrating optical components such as phase contrast spotlights and objective lenses. These components work together to accurately illuminate the observed cell sample and collect reflected or transmitted light to form clear images. At the same time, combined with a high-sensitivity camera, optical signals can be converted into digital images, supporting multiple imaging modes such as bright field, fluorescence, and relief effects, thus meeting different experimental needs.
2. Fluorescent labeling and dynamic observation: By using specific fluorescent dyes, specific proteins or organelles in cells can be labeled. With the help of fluorescence microscopy technology, the activity of cells can be transformed from a static "snapshot" to a dynamic "movie", displaying the internal activity of cells in real time, such as molecular transport, interactions between organs, etc.
3. Automation control and environmental maintenance: It is also equipped with an automation control system that can accurately adjust environmental parameters such as temperature, humidity, and carbon dioxide concentration, providing a stable and suitable growth environment for cells. This not only ensures the activity of cells during the experimental process, but also reduces interference caused by changes in environmental factors, improving the accuracy and reliability of the experiment.
Precautions for using cell environment imaging system:
1. Mercury lamp usage guidelines: The mercury lamp switch should not be repeatedly turned on frequently. The interval between two switches must be at least 30 minutes, otherwise it may affect the lifespan and performance of the mercury lamp.
2. Environmental temperature control: The microscope room should maintain a low temperature environment. A stable temperature helps maintain instrument accuracy and cell state, reducing errors caused by temperature fluctuations.
3. Light avoidance and earthquake prevention measures: During the experiment, attention should be paid to avoiding light to prevent damage to cells or interference with experimental results; At the same time, avoid vibration as it may cause image blurring, focus shift, and other issues that affect imaging quality.
4. Key points for selecting culture medium: When preparing samples, the culture medium formula used should not only contain appropriate nutrients to promote cell growth and development, but also consider factors such as pH value, buffering capacity, and osmotic pressure. Inappropriate culture medium can affect cell expression, phenotype, and appearance behavior. It is recommended to use phenol red free culture medium and reduce serum concentration to decrease background signal.
5. Requirements for aseptic operation: During the process of collecting cells and other operations, aseptic operation must be strictly followed to prevent contamination. Foreign particles or cell debris can interfere with image quality and affect experimental accuracy.
6. Necessity of probe optimization: Before conducting formal experiments, the concentration and incubation time of fluorescent probes should be optimized. It is necessary to ensure that the cell viability is not affected after incubation, while also ensuring that appropriate fluorescence signal intensity can be detected. In addition, potential background fluorescence can be reduced by adding a washing step.
7. Stable environmental conditions: Throughout the imaging experiment, strictly control and maintain stable environmental conditions, including temperature, humidity, and gases (oxygen and carbon dioxide). A stable environment not only maintains cellular health, but also reduces the risk of focus drift. For example, before configuring the acquisition settings, ensure that the microplate is located on the instrument table or in the environmental control room for thermal balance; If a medium is added to the hole during the delay experiment, it must be ensured that it is at the same temperature as the medium inside the imaging container.
8. Application of objective lens heating module: For experiments using water or oil mirrors for imaging, due to the contact between the objective lens and the sample through the medium, the temperature of the objective lens will have a significant impact on the sample. At this time, an objective lens heating module needs to be equipped to ensure that the sample is in a suitable temperature environment.
9. Utilization of focal plane control system: Fully utilize software autofocus functions (such as best focus), focal plane stabilization system (AFC), and accurate focus (closed-loop) to ensure clear images can be obtained at different time points, improving imaging quality and data accuracy.
10. Advantages of remote monitoring: If the device has remote monitoring function, it can be reasonably used to monitor the progress of the experiment in real time, discover and solve problems in a timely manner, and improve the efficiency and success rate of the experiment.