Microbial inoculation operation

Microbial inoculation operation

1. Slant inoculation (with Staphylococcus aureus)
(1) Before operation, wipe your hands with 75% alcohol, and light the alcohol lamp after the alcohol evaporates.
(2) Hold the strain tube and the inclined plane between the thumb of the left hand and the other four fingers, so that the inclined plane and the side with strains are upward and in a horizontal position.
(3) Rotate the strain and the tampon on the inclined plane first, so that it can be easily pulled out during inoculation.
(4) Hold the inoculation ring in the left hand (like holding a pen), and sterilize the ring end with a flame, and then sterilize the rest of the test tube that may extend into the test tube.
(5) Use the ring finger, little finger and palm of the right hand to pull out the strain tube and the cotton plug or test tube cap of the inclined test tube to be connected at the same time, and then let the test tube mouth be slowly overheated to sterilize (do not burn it too hot).
(6) Extend the burnt inoculation loop into the bacterial culture tube, touch the inoculation loop on the inner wall of the test tube or the medium without bacterial moss, let it cool down, and then gently scrape a little bacterial moss, and then remove the bacteria from the bacteria. Pull out the inoculation loop from the seed tube.
(7) Quickly extend the inoculation loop stained with the strain into another inclined test tube to be connected. From the bottom of the bevel upwards, make a dense line in a “Z” shape back and forth. Sometimes the inoculation needle can also be used to draw a line in the center of the medium for slant inoculation, so as to observe the growth characteristics of the strain. .
(7) Quickly extend the inoculation loop stained with the strain into another inclined test tube to be connected. From the bottom of the bevel upwards, make a dense line in a “Z” shape back and forth. Sometimes the inoculation needle can also be used to draw a line in the center of the medium for slant inoculation, so as to observe the growth characteristics of the strain.
(8) After the inoculation is completed, take out the inoculation ring to burn the mouth of the tube and plug it with a cotton plug.
(9) Sterilize the inoculating ring by burning it in red. Put down the inoculating loop and tighten the cotton plug.
2. Liquid inoculation
(1) The slanted medium is connected to the liquid medium. This method is used to observe the growth characteristics of bacteria and the determination of biochemical reactions. The operation method is the same as before, but the mouth of the test tube is inclined upward to prevent the culture liquid from flowing out after the bacteria are inserted. , rub the inoculation ring and the inner wall of the tube a few times to wash the bacteria on the lower ring. After inoculation, plug the cotton plug and tap the test tube gently in the palm of the hand to fully disperse the bacteria.
(2) Inoculate the liquid medium from the liquid medium. When the strain is liquid, use a sterile pipette or dropper for the joint in addition to the inoculation loop. When inoculating, just pull out the cotton plug beside the flame, pass the nozzle through the flame, suck the bacterial liquid into the culture solution with a sterile pipette, and shake well.
3. Plate vaccination
Bacteria were streaked and spread on plates.
(1) Inoculation by streaking See separation streak method.
(2) Coating and inoculation After sucking the bacterial solution into the plate with a sterile pipette, evenly spread it on the surface of the plate with a sterilized glass rod.
4. Puncture inoculation
The strains are inoculated into solid deep medium. This method is used for inoculation of anaerobic bacteria or for observation of physiological properties when identifying bacteria.
(1) The operation method is the same as above, but the inoculation needle used should be straight.
(2) Pierce the inoculation needle from the center of the culture medium until it is close to the bottom of the tube, but do not penetrate it, and then slowly pull it out through the original piercing method.