What is the theory of an anaerobic experiment?
An anaerobic culture experiment is an experiment that uses containers such as an anaerobic incubator to create an anaerobic or microaerobic environment and then simulates the environmental temperature and other conditions required for the growth of anaerobic bacteria. The experimental process of anaerobic culture is removing oxygen, gradually affecting the environment, maintaining the anaerobic and gas temperature environment, cultivating bacteria, and then conducting experiments to conclude.
The following is the whole experiment procedure and theory:
1. First, we use a vacuum pump to make the anaerobic incubator inner in a vacuum state.
2. Make the anaerobic incubator filled with the mixture gas of either
(1).80%N2 +10%CO2+10% H2 or
(2).85% N2 +10% CO2 +5% H2
Here we have added H2 to react with trace oxygen in the anaerobic incubator.
3. We use a Palladium particle molecular sieve(about 500g) as the catalyzer to make hydrogen react with oxygen.
2H2+O2--(Palladium particle molecular sieve)-->2H2O
*Precautions: catalyst palladium particles will lose activity due to hydrogen sulfide or other volatile metabolites produced by bacteria. Experience shows that the service life of the catalyst is generally about 100 cycles. Many factors can affect the service life of the catalyst. If the catalyst is affected by moisture, it will lose its activity partially or completely. Therefore, it must be completely dried before use. Please dry the catalyst palladium particles and store them in a closed, dry, and clean environment at room temperature.
4. Verify the inner anaerobic status
The oxygen indication paper can indicate the anaerobic status.
The oxygen indication paper has the following performance:
(1). Reversibility of color change: The color of the oxygen indicator can change reversibly from blue to red and from red to blue many times.
(2). Reaction time of discoloration: when the oxygen indicator is used together with the deaerator, the oxygen indicator turns red within 4 hours after the oxygen concentration drops to 0.1%; When external oxygen enters, the oxygen indicator turns blue within 30 minutes. (at 25 ℃)
(3). Indicated oxygen concentration range: when the oxygen concentration in the sealed container is below 0.1%, the oxygen indicator is red; When the oxygen concentration in the sealed container is above 0.5%, the oxygen indicator is blue.
How to use the oxygen indication paper?
(1). In the absence of oxygen, the red oxygen indicator turns blue within 30 minutes after being taken out of the outer packaging bag and can be used safely. After unpacking the outer packaging bag, remove the required oxygen indicator, and the remaining part shall be immediately deaerated and sealed with a deaerator.
(2). When used in combination with the deoxidizer to seal the articles, the deoxidizer, oxygen indicator, and the sealed articles can be sealed in the container at the same time, and the indicator can be fixed at a position convenient for observation.
(3). Do not take the indicator out of the inner packaging bag for naked use to avoid contaminating the sealed articles or eating by mistake. In case of eating by error, the oxygen indicator is non-toxic, so it is harmless to the human body.
(4). The temperature of the oxygen indicator is usually 2 ~ 40 ℃, and the relative temperature of the environment is rh30% ~ 100%. The oxygen indicator shall be stored without oxygen at a low temperature (2-10 ℃) and away from light. We must try to avoid storing or using it under direct sunlight.
(5). The oxygen indicator can be used to indicate the presence or absence of oxygen, but it cannot be used for the environment of gas and heat generation.
5. Under the confirmed oxygen-free environment, the required gas can be gradually replaced according to the needs of the experiment, or no need to replace the inner gas. Just maintain the internal environment.
During the anaerobic culture experiment, except for the anaerobic incubator, we also need the following supporting items:
1. Vacuum pump and vacuum valve.
2. Palladium particle molecular sieve(about 500g) and desiccant for the palladium particle molecular sieve.
3. Oxygen indication paper.
4. The special gloves.
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