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What should be paid attention to when using living cell imaging technology

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What should be paid attention to when using living cell imaging technology

Date of release:2020-08-22 Author: Click:

In recent years, the imaging technology of living cells has been continuously improved, and many cells have been found, which is mainly due to the poor performance of fluorescent labeling technology, electronic technology and data processing. The most obvious advantage of Living cells is that they can record the cells and the dynamic process of the whole organism. Many processes can be visualized and more convenient for recording. In the use of cell imaging technology, we also need to pay attention to the following key points:

Living cell imaging technology

1. Correct culture conditions

Appropriate cell culture medium (including buffer and growth serum), appropriate culture vessels and aseptic technology. Constant temperature 37 ° C. To neutralize acidification caused by metabolites, a bicarbonate (HCO3 -) and dissolved carbon dioxide (CO2) buffer system was used to maintain pH 7.4. If CO2 is not available, a buffer containing 10-20 mm HEPES can also be used. Phenol red is a common pH monitoring method. The choice of culture containers is also important. Cells usually grow better on plastic, but glass provides higher optical quality. Glass, on the other hand, usually requires a coating, such as polylysine or polyornithine. Bacterial contamination is always a problem, so the use of antibiotics is common, and if a particular protocol requires the avoidance of antibiotics, more care should be taken in the preparation and maintenance of the culture.

Living cell imaging requirements

2. Protect cells from phototoxicity

Fluorescence imaging may have potential harmful effects on cell homeostasis due to phototoxicity, which may come from different sources. Organic molecules in cells, such as porphyrins and flavins, will absorb light and degrade when they react with oxygen, thus releasing reactive oxygen species, such as superoxide radical, hydroxyl radical and hydrogen peroxide, which will cause cell damage. Fluorescent dyes themselves can also produce free radicals because they can react with oxygen in the excited state. Phototoxicity can be avoided by some simple and effective measures.

3. Appropriate frame rate

When observing changing activities, researchers need to ensure that the image frame rate is appropriate for tracking these events. A low frame rate means a loss of time resolution and a poor representation of the process of interest in the log file. On the other hand, too high frame rate will increase the risk of phototoxicity, resulting in larger files. However, if the activity to be recorded is so fast that the required frame rate leads to phototoxicity, in this case, it is very helpful to improve the sensitivity of the camera and achieve satisfactory results with the required frame rate at a lower excitation light intensity and lower gain.

Characteristics of living cell imaging

In fact, the use of living cell imaging technology can provide evidence, and many biological problems can be judged by Immune tumor microenvironment. We have cell molecules large and small, and specific imaging systems can also be used to observe molecules in moving animals. If you need this, please feel free to contact us.

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Key word:Livingcellimagingtechnology,Livingcellimagingrequirements,Characteristicsoflivingcellimaging

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Living cells