Sample Addition and Troubleshooting Methods for Cell Culture Plate

Cell culture plate follows strict aseptic principles when performing cell operations, and all operations should be standardized and scientific to avoid causing additional effects on cell growth. One of the most common issues is how to ensure uniformity in cells after adding samples and minimize the impact of medium changes on cell growth.

Q: The lids of the 96- and 24-well plates or culture dishes are very loose, which facilitates ventilation, but will contaminants such as bacteria and mold also enter?

A: Loose lids are classified as semi-open culture, and the goal is to facilitate ventilation (in fact, to allow CO2 outside the culture dish to fully exchange with the culture dish to maintain the pH of the medium).

For everything that is advantageous, there will be disadvantages, which of course will increase the possibility of contamination. In addition, this will cause the liquid inside the culture dish to evaporate, which is worth noting for precise dosage of drugs. Therefore, the following two measures are required: a. air in the incubator must be clean (regularly exposed to ultraviolet light, alcohol scrubbed, and the incubator should be switched off as little as possible) b. humidity in the incubator must always be maintained at 100% (placing sterile distilled water in the container in the incubator).

Just like culture dishes, they are also inverted containers with lids and will not contaminate. This is mainly because the "L"-shaped edges of the lid generate negative pressure airflow, and microorganisms only adhere to the dust on top, which cannot pass through the negative-pressure side edges caused by air flow. The ventilation effect is only achieved through the diffusion of air, and it will not transport bacteria. Gondong is a professional manufacturer in this, if you are interested in oem cell culture plate, you can contact us for more information.

Q: When using a 24-well plate, some wells are manipulated (in a super clean bench), and other wells still need to be cultured. I am worried that it will cause contamination. What should I pay attention to?

A: If the operation inside the super clean bench is standardized, it should be okay. Make full use of the lid of the cell culture plate, and try to expose only the wells to be treated, and cover the other wells with the lid.

Before use, consider using all wells to the fullest extent. If only a few wells are needed, use only one side and cover the others with a lid. Get used to using the wells on the right side first (right-handed for easy sampling).

During the operation, use several slides to raise one side of the plate, do not fully open the lid, and generally there should be no problem.

Q: The cells in the cell-seeded culture plate always aggregate in the peripheral part. How should I deal with it?

A: As a professional medical oem supplier, we would ask you first: How do you mix your cells? Do you use pipetting or shaking the culture plate? If it is the latter, and it is shaken in circles, it is very likely that the cells are thrown to the periphery due to the centrifugal force, resulting in less cells in the middle and more at the periphery!

Here is a good way: before culturing the cell-seeded plate, put the cell culture plate in the incubator for several hours to saturate it, and be gentle when seeding the cells. Slowly add the cell suspension into the wells of the culture plate using a dropper on the side of the holes, and the cells will grow evenly. Remember not to use an oscillator, otherwise your cells will aggregate as you described.

The smaller the diameter of the holes of the cell culture plate, the more obvious this phenomenon. For 24- and 96-well plates, this phenomenon is inevitable, because the medium adheres to the wall, so the medium in the holes is not a flat surface, but is higher at the edge, like a concave mirror. Using these two types of plates, because of the need for intervention, the amount of culture medium cannot be sufficient, causing "edge aggregation" phenomenon to occur when the cells grow with the medium. If you increase the amount of seeded cells to enable the central part to have an even distribution of cells, it depends on what you need to observe. If it is MTT or immunohistochemistry, the results will be affected because the cells are layered.

When digesting the cells, pay attention to uniform pipetting to avoid cell aggregation, and ensure that the amount of medium in the wells is sufficient. When seeding, add enough culture medium, and when intervening, replace the medium. The amount of intervention medium added is equal to the amount of culture medium during seeding, and this will improve the phenomenon of "edge aggregation." It's worth a try.