Preparation and observation of water-soaked specimen slices of mycobacteria
Preparation and observation of water-soaked specimen slices of mycobacteria
The preparation and observation of mold water-impregnated specimen slices are mainly applied to observe mold morphology.
Operation method
Preparation and observation of water-soaked specimen slices of mycobacteria
Principle
The trophozoites of molds are branched filaments. They are much larger than bacteria and actinomycetes and are divided into basidiomycetes and aerial mycelium. The aerial mycelium can be divided into reproductive filaments. The reproductive filaments of different molds can form different spores. Mold mycelium is thicker, the cell is easy to shrink and deformation, and the spores are easy to fly apart, so the production of specimens commonly used lactic acid carbonate cotton blue staining solution. This staining solution made of mold specimens are characterized by: the cells are not deformed, with bactericidal antiseptic effect, and is not easy to dry, can be maintained for a longer period of time, the solution itself is blue, there is a certain staining effect. The use of mold cultured on cellophane as observation material, you can get clear, complete, maintain the natural state of the mold morphology; can also be directly picked in the plate of mold growth in water immersion film observation.
Materials and Instruments
Rhizoctonia Aspergillus Penicillium Move 1. Preparation of water-immersion slide observation method Add a drop of lactic carbolic acid cotton blue staining solution or distilled water on the slide, use a dissecting needle to pick a small amount of mold mycelium with spores from the plate with mold growth, moistened with 50% ethanol, and then wash the immersed mycelium with distilled water, and then put it into the droplet on the slide, and carefully use the dissecting needle to disperse the mycelium. Cover the coverslip (do not make bubbles, and do not move the coverslip again), first with low magnification, if necessary, switch to high magnification microscopy and record the observations. 2. cellophane dialysis culture observation method (1) Selection and treatment of cellophane Select cellophane that allows nutrients to pass through. You can also collect the cellophane used for commodity packaging, boil it with water, and then rinse it with cold water. If the cellophane becomes hard after this treatment, it must be unusable, and only those that are soft can be used. Those available cellophane cut into the appropriate size, soaked in water, sandwiched between the old newspaper, and then together in a petri dish 121 ℃ sterilized for 30min standby. (2) the culture of strains of bacteria according to the aseptic method, pour the plate, condensation, with sterilized tweezers clip the sterile cellophane attached to the plate, and then use the inoculation ring to dip a little mold spores, in the cellophane above the paper gently shake down. Then put the plate at 28-30 ℃ for 3-5 days, Aspergillus and Penicillium can grow a single colony on the cellophane (strong aerial root molds, the formation of colonies spread throughout the plate). (3) Preparation and observation Cut a small piece of cellophane with mycelium and spores after 3-4 days of cellophane dialysis culture, first put in 50% ethanol immersion, wash off the spores, and drive away the air bubbles on the bacterium, and then attached to a clean slide face up, add 1-2 drops of lactic acid carbonate cotton blue solution, carefully cover the coverslip (pay attention to do not produce bubbles), and do not move the coverslip, so as not to mess up the mycelium, and not to move the coverslip, so as not to mess up the mycelium. Do not move the coverslip to avoid messing up the mycelium. After the specimen is ready, use low magnification to observe first, and then change to high magnification if necessary. Observe the mycelium with or without septa, pseudoroots, foot cells and other special forms of mycelium. Pay attention to the shape and structure of the asexual reproductive organs, the way of spore implantation and the shape and size of the spores. Caveat 1. cover the coverslip carefully (be careful not to create air bubbles), and do not move the coverslip to avoid messing up the mycelium. 2. When using a microscope to observe the specimen, use low magnification to observe first, and then change to high magnification if necessary. Pay attention to the observation of mycelium with or without septa, with or without pseudoroots, foot cells and other special forms of mycelium. 3. Record the morphological changes of mold in time. Common Problems 1. rhizobium is highly aerial, forming colonies spread over the whole plate. 2. Lactic acid carbolic acid cotton orchid staining solution, the main ① can make the cells not deformed ② antiseptic and bactericidal effect ③ is not easy to dry, can be stored for a long time ④ the solution itself is blue in color, there is a certain staining effect. For more product details, please visit Aladdin Scientific website.
Cotton Blue Staining Solution Lactic Carbonate 50% Alcohol
Scissors, slide, tweezers, dissecting needle, microscope.