First, the tomb principle The colony morphology refers to the formation of a certain microbe on a certain medium formed by a single cell. Bacteria, actinomycetes, yeasts and molds, the colonies formed by each type of microorganism under certain culture conditions have certain relative characteristics, and these characteristics are observed to distinguish the major microorganisms and to initially identify and identify microorganisms. Simple and fast, it is often used in research and production practice.
The blood medium is rich in nutrients and viscosity to culture the bacteria. The colonies grown on the medium are highly viscous when fixed, and can be firmly attached to the slide or cover glass for tableting and observation. In addition, the blood medium is a better medium for preserving the strain, so that the colony produced by the medium can be stored for a long time.
Second, the equipment round brown nitrogen-fixing bacteria, Bacillus subtilis, Streptomyces griseus, Saccharomyces cerevisiae, Penicillium single colony plate, Staphylococcus albus;
Bouin's fixative, 0.01% crystal violet solution, sheep blood or rabbit blood, meat paste peptone agar medium, alcohol, etc.
Third, the operation steps 1. The individual colony characteristics of B. aureus, Bacillus subtilis, Streptomyces griseus, Saccharomyces cerevisiae, Penicillium were observed, and the results were recorded according to the method described in the experimental one colony.
2. Preparation of blood agar medium:
(1) Component pH 7.6 meat paste peptone agar medium 10 ml, sterile defibrinated sheep blood (or rabbit blood) 1 ml.
(2) The meat paste peptone agar medium is heated and dissolved.
(3) When it is cooled to 45 ° C, add 1 ml of sterile defibrinated sheep blood to the culture medium aseptically, and immediately shake to mix the blood and the agar medium thoroughly.
(4) Quickly pour into the plate aseptically to form a blood agar plate, taking care not to create bubbles.
(5) Set at 37 ° C overnight, aseptic growth can be used.
(6) Take out the sterile plate and perform single-column isolation by streaking. Because the surface of the agar is to be kept intact, the round head smooth glass rod should be used for inoculation. Be careful not to cut the agar. After inoculation, the cells were cultured in a greenhouse at 37 ° C, and the results were observed the next day. If a single colony appears, the next experiment can be performed.
3. Bacterial colony preparation (1) Bacterial colony printing Select a smaller single colony, cut a square agar about 10-15 mm around the colon with a knife, and place the colony side facing down on the agar block. On the coverslip, then carefully place the coverslip in Bouin's fixative solution with agar block for about 1 hour until the agar is completely white and remove it. Carefully peel off the agar. Be careful not to damage the colonies when peeling, only colonies Leave and attach to the coverslip.
(2) Gently rinse the coverslips with colonies with a stream of fine water.
(3) Dyeing with 0.01% crystal violet dye solution for 3 minutes, washing with water, and blotting.
(4) Dehydration Place the coverslips in low to high concentrations of alcohol (35%, 50%, 60%, 70%, 80%, 95%, 100%) for 5 minutes each, but at 100% alcohol Immerse for 15 minutes.
(5) The dehydrated coverslip was taken out transparently and placed in xylene for 2 minutes.
(6) Sealing The coverslip was taken out from the xylene, and the xylene around the colonies was wiped off with a soft paper. Immediately add a small drop of Canadian latex to the center of a clean glass slide, quickly reverse the coverslip and gently place it on the slide to allow the Canadian latex to spread over the coverslip, taking care not to create bubbles, if there are bubbles Gently press out. The colony characteristics were observed with a low power microscope. Wait until the latex is completely dry, and store it in a wooden spoon one or two days later.
Fourth, the experimental report
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