What should be paid attention to in microbial inoculation and how to carry out microbial inoculation

It depends on whether you are inoculated with the fungus.

Bacteria or actinomycetes and other types have their own particulars, but the common points are as follows:

1.Rational selection of culture medium.

Different strains have different optimal mediums, such as Gao's.

culture medium, beef peptone, PDA, etc.;

2.Pay attention to proficient inoculation, pay attention to aseptic operation, keep the table ultra-clean, and reduce the probability of miscellaneous bacterial infection;

3.Adjust the culture environment, different microorganisms such as fungi, bacteria or actinomycetes have different requirements for temperature and humidity.

4. Pay attention to the incubation time, microorganisms have their reasonable incubation time, too short is not conducive to activation, too long is easy to be infected.

1) Inclined inoculation: A small amount of strains are selected from the inclined plane of the grown microorganisms and inoculated on another blank inclined medium.

1) Before inoculation, use a marker pen to indicate the name of the bacteria and the date of inoculation at a distance of 2 3cm from the mouth of the test tube.

2) Place the tube in the palm of your hand and hold it between your fingers so that the two tubes are in a "V" shape.

3) Use the flame outer flame to burn the inoculation ring and needle red, and then pass the inoculation ring back and forth through the flame several times.

4) Remove the tube plug with your little finger, ring finger and palm and hold it in your hand.

5) Slightly burn the mouth of the test tube on the flame for a week.

6) Extend the burned inoculation ring into the strain tube, first touch the medium part without growing bacteria to cool it so as not to burn the bacteria, and then gently touch the fungus with the ring, scrape off a little culture, and slowly pull the inoculation ring out of the test tube, and quickly extend it into another test tube, and draw a line (wave or straight line) on the inclined plane, so that the bacteria stick to the medium.

7) Draw out the inoculation loop and cauterize the nozzle.

8) Plug in the tampon.

9) The inoculation ring is sterilized by flame burning.

2) Liquid inoculation: inoculated by inclined strains in liquid medium.

1) Before inoculation, use a marker pen to indicate the name of the bacteria and the date of inoculation at a distance of 2 3cm from the mouth of the test tube.

2) Place the tube in the palm of your hand and hold it between your fingers so that the two tubes are in a "V" shape.

3) Burn the inoculation loop red with a flame and pass the inoculation loop back and forth through the flame several times.

4) Remove the tube plug with your little finger, ring finger and palm and hold it in your hand.

5) Slightly burn the mouth of the test tube on the flame for a week.

6) Extract the inoculation loop and cauterize the nozzle.

7) Plug in a tampon.

8) The inoculation ring is sterilized by flame burning. It should be noted that my personal opinion is that the sterile environment, speed, and proficiency, the liquid is relatively simple, and the bevel should ensure the purity of the source.

The following points should be noted for microbial inoculation:

Reasonable selection of culture medium, different strains have different optimal mediums, such as Gao's medium, beef egg finger head white peptone, PDA, etc.; Pay attention to proficient inoculation, pay attention to aseptic operation, keep the table ultra-clean, and reduce the probability of miscellaneous bacterial infection; Adjust the culture environment, different microorganisms such as fungi, bacteria or actinomycetes have different requirements for temperature and humidity. Pay attention to the incubation time, microorganisms have their reasonable incubation time, too short is not conducive to activation, too long is susceptible to infection.

The common methods of inoculation are: plate marking inoculation, inclined plane inoculation, pouring culture method, puncture inoculation method, and liquid inoculation method.

1. Plate scribing separation method.

Plate scribing and separation: It refers to the mixing of microorganisms or different cells in the same microbial population on the surface of the plate medium with inoculation rings, and more independently distributed single cells are obtained by partitioning and line dilution, which grow and multiply into a single colony after culture, and this single colony is usually regarded as a pure breed of microorganisms to be isolated.

2. Inclined plane inoculation method.

This method is mainly used for the pure culture of a single colony, to preserve the culture, or to observe certain characteristics of the bacteria.

3. Puncture inoculation method.

Piercing blind thorn culture of cracked antch liquid refers to a method of inserting the inoculation needle with the microorganisms to be cultivated into the semi-solid medium (agar content) and carrying out the deep culture of microorganisms in solids, mainly used for the cultivation of anaerobic or facultative anaerobic microorganisms.

Fourth, liquid inoculation method.

Liquid culture refers to a culture method in which microorganisms are directly inoculated in a liquid culture medium and continuously shaken or stirred to make microorganisms grow and multiply uniformly in the liquid culture medium. Liquid culture is suitable for aerobic microorganisms and plant tissue culture, with the aim of rapidly obtaining a large number of propagules.

5. Coating inoculation method.

A method of operation in microbiology experiments. Because adding the bacteria-containing material to the hot medium and then pouring the plate is easy to cause the death of some heat-sensitive bacteria, and the use of dilution and inverted platform method will also make some strict aerobic bacteria affect their growth due to the lack of oxygen immobilized in the middle of the agar, so the more commonly used method of pure species isolation in microbiology research is the coating plate method.

<> plate scribing inoculation method is a common technique for isolating and culturing bacteria, with the aim of forming a single colony or isolating a single strain from a culture mixed with multiple bacteria for easy identification and identification. Inclined inoculation method:

It is mainly used to transplant pure bacteria and proliferate them for identification or preservation of strains. Pouring culture method contains files: mainly used for counting bacteria and molds in feed.

Puncture inoculation method: It is mostly used for stuffy inoculation of disaccharide, semi-solid, gelatin and other media, and the method is similar to that of inclined inoculation.

Microbial inoculation is the most commonly used basic operation in microbiology research, mainly for the isolation and purification of microorganisms. This technology is to pick the required microorganisms from one petri dish with special tools such as inoculation loops or inoculation needles under sterile conditions, and transfer them to another medium for culture, so as to achieve the purification and identification of the required microorganisms and obtain simple colonies without bacterial contamination.

There are several main methods:

1. Inoculation ring or inoculation needle: The ear is mainly used to connect from the inclined seed to the inclined plane or seed liquid;

2. Pipette or pipette: mainly used to change from seed liquid inoculation to seed liquid;

3. Suction filter bottle: mainly used to inoculate the seed liquid from the shake flask into the seed tank or fermentation tank.