Cell culture basic knowledge (machine) – 2

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Hello! Today, I’m here as the second content of the cell culture basic knowledge device.

As I told you last time, today I will give you information related to the incubator and microscope.

 

 

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  CO2 Incubator

When culturing cells in a laboratory, they are cultured by putting cells in a CO2 incubator. The incubator must maintain the optimal environment by controlling a certain temperature and CO2 for cell culture. The most commonly used incubator in the experiment uses a CO2 incubator with a CO2 adjusted from 5% to 10%.

When media containing phenol red components that can be checked for pH is used in the CO2 incubator, the color of the media changes according to the pH.
If it’s acidic, it turns orange, if it’s alkaline, it turns orange.
Depending on the concentration of CO2, the pH of the medium can be affected.

In general, sodium bicarbonate is included in the media, which acts as a buffer and interacts with CO2 to properly adjust the pH of the media. It also maintains osmotic pressure and provides energy sources.
However, sodium hydrogen carbonate does not have an optimal buffering effect in the pH range of 7 to 7.5 at 37°C and requires a special environment of 5 to 10% CO2/90 to 95% air.
Therefore, when culturing cells using media containing sodium hydrogen carbonate, the pH of the media should be adjusted using CO2 incubator.

If sodium hydrogen carbonate needs to be replaced, HEPES buffer is also used, which has 37℃ dptj pKa7.31 and does not pass through the cell membrane, making it ideal for cell culture media.
However, in the case of HEPES buffer, when used in addition to media, the concentration of NaCl must be lowered to match the osmotic pressure.

In addition, since cells are very sensitive to temperature and humidity, the air circulation inside the incubator should be good, so that the difference in temperature, humidity, and CO2 by region should not be large.

The inside of the incubator should be disinfected and cleaned regularly to minimize contamination.

 

 

 

 

 

 

 

 

 

 

 

• An inverted microscope

You need a microscope to see cells that you can’t see with your eyes, but it’s essential in cell culture.

Among many types of microscopes, provincial microscopes are usually used.

Contrary to a normal microscope, a microscope is designed to irradiate light downward from a light source at the top of the specimen table, capture the image with an objective lens from the bottom, and refract it by a prism so that it can be observed in the front eyepiece. It is said that the shape and function of culture cells attached to the bottom of the container and proliferating can be observed alive.

[Naver Encyclopedia of Knowledge] An inverted microscope [Inverted microscope, 倒顯微鏡微] (Life Science Dictionary, first edition 2008, revised edition 2014, Kang Young-hee)

Simply put, the other microscope is a microscope that is often used to observe cells in a cell-culturing laboratory where the lens is attached to the sacrificial table while the lens is attached to the sacrificial table.

Cell culture-related tasks are different depending on the condition of the cells by observing them directly with a provincial microscope every day, so you should control the magnification with the provincial microscope and carefully observe the cells.

My eyes are a little bad, so I lower the brightness a little bit and make observations, but I think it’s easier to check the cell status by raising the brightness.
However, I use it to reduce eye fatigue by lowering the brightness a little because I may have to look at the microscope for a few hours.

The microscope management method has a long life of the lamp, so make sure to turn off the power if you don’t use it for a long time or when you leave work, cover it to block the light.
Eye contact lenses should also be cleaned from time to time to minimize mis-checking of cell conditions due to dust on the lenses.

Today, I explained about CO2 incubator and provincial microscope.

We will give you more device information, reagents and know-how related to cell culture, so please refer to it!!

Thank you!

– Note : Basic knowledge of cell culture (machine) – 1