Bacteria isolation and inoculation method
Bacterial isolation and inoculation is an important step in artificial isolation and culture of bacteria. At the same time, it is of great significance in the research of bacteriology, the preparation of biological products such as vaccines, toxoids, antitoxins and immune serum, genetic engineering and industrial and agricultural production.
The inoculation method of bacteria can be divided into line drawing method, puncture culture method, slant inoculation method, liquid medium inoculation method, etc. according to the purpose and the medium. Pay attention to aseptic operation when inoculating to avoid contamination.
1. Plate line drawing method: There are many methods of line drawing inoculation, such as parallel line drawing method, partition line drawing method, checkerboard drawing method, etc. The tested specimens, such as pus, feces, urine, cerebrospinal fluid, etc., are often mixed with a variety of bacteria. Through the line drawing method, the mixed bacteria can be dispersed and grown on the surface of the agar plate, each forming a different colony, and then according to the morphological characteristics of the colony. , select a single colony and continue to cultivate, so as to obtain the pure breed of bacteria for further research, identification or preservation. Methods as below:
(1) The inoculation ring is sterilized by burning on a flame, and after cooling, take a few specimens, and draw a continuous parallel line in one area on the solid agar plate, which accounts for about 1/10 of the area of the plate, and is the first group of lines.
(2) Sterilize the inoculation loop by cautery, and after cooling, turn the petri dish about 60°, draw the inoculation loop through the first set of lines and then draw the second set of lines, and carry out the third and fourth sets of line drawing inoculation in the same way.
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2. Slant medium inoculation method: Slant medium is often used for the propagation of pure strains, as a short-term preservation of strains or to observe certain biochemical properties, such as agar slants, citrate slants, etc. After inoculation of the solid slant, a single and neat bacterial lawn grows along the inoculation line and cannot be divided into single colonies. Methods as below:
(1) The inoculation ring is sterilized by burning, and after cooling, dip a little bacterial moss.
(2) The slant culture medium is slanted upward, the nozzle is sterilized by flame cautery, the inoculation ring is inserted, and a straight line is drawn along the midline of the slant from bottom to top, and then a continuous serpentine line is drawn from the bottom to the top.
3. Puncture inoculation method: The puncture inoculation method is mostly used to observe the growth of bacteria in semi-solid medium and some biochemical reactions. The medium inoculated by puncture method includes semi-solid agar medium, lead acetate medium, gelatin medium and so on. The inoculation method is as follows:
(1) After the inoculation needle is sterilized by burning on the flame, take a little bacterial moss after cooling.
(2) After the nozzle of the medium is sterilized by flame cauterization, the inoculation needle is vertically pierced into the center of the medium to a depth of 0.5cm near the bottom, and then exit along the original route.
After incubation after inoculation, the motile bacteria spread out along the inoculation line and grow like a brush; the non-motile bacteria did not spread.
4. Liquid medium inoculation method: Broth medium, peptone water and various monosaccharide fermentation tubes and other liquid medium are all inoculated by this method. After inoculation, the different growth conditions of bacteria can be observed by culturing, and the determination of biochemical characteristics can also be carried out. Methods as below:
(1) The inoculation ring is sterilized by burning, and after cooling, dip a little bacterial moss.
(2) After the nozzle of the liquid medium is sterilized by flame burning, insert the inoculation ring, grind it gently on the tube wall close to the liquid surface, and dip it in a small amount of liquid to mix the bacteria in the liquid medium.
Post time: Mar-21-2022