Mold mycelium is thicker, the cells are easy to contraction deformation, and spores are easy to fly apart, so the specimen is commonly used lactic acid carbonate cotton blue staining solution. This staining solution made of mold specimens are characterized by: (a) the cells are not deformed; (b) with bactericidal antiseptic effect, and is not easy to dry, can be maintained for a longer period of time; (c) the solution itself is blue, there is a certain staining effect.
Operation method
Morphological observation of molds
Principle
The morphology of mold in its natural state of growth is commonly observed on slides, which is inoculated with mold spores on a suitable medium on the slide, and then observed with a microscope after cultivation. In addition, in order to get clear, complete, maintain the natural state of the mold form can also use cellophane dialysis culture method for observation. This method is the use of cellophane semi-permeable film properties and light transmission, the mold growth in the agar medium covering the surface of the cellophane, and then the long bacterial cellophane cut a small piece of cellophane, placed on the slide with a microscope to observe.
Materials and Instruments
Aspergillus, Penicillium, Rhizoctonia, Trichoderma Move 1. General observation methodOn a clean slide, drop a drop of lactic acid, carbolic acid cotton blue staining solution, with a dissecting needle from the edge of the mold colony to take a small amount of mycelium with spores placed in the staining solution, and then carefully pick the mycelium scattered, and then carefully covered with a coverslip, pay attention to do not produce air bubbles. Place under the microscope and observe with low magnification first, then change to high magnification if necessary. For more product details, please visit Aladdin Scientific website.
Lactic acid, phenolic acid, cotton blue staining solution Glycerol Chartreuse medium plate Potato medium
Slides Coverslips U-shaped rods Scalpels Cellophane Filter paper
2. Slide observation
(1) Put the filter paper slightly smaller than the inner diameter of the bottom of the petri dish into the dish, and then put a U-shaped slide, on which a clean slide is placed, then two coverslips are placed diagonally at the ends of the slides, cover the lid of the dish, and then stack several sets of petri dishes (depending on the needs of the device) so that the device is wrapped up, and then sterilized at 1.05 kg/cm2, 121.3 ℃ for 20 minutes or dry heat sterilization, and then standby.
(2) Pour 6~7 ml of sterilized potato dextrose medium into a 9 cm diameter sterilized Petri dish, and after solidification, use a sterile scalpel to cut the agar block into 0.5~1 cm2 pieces, and use the tip of the knife to shovel up the agar block and place it on the slide in the sterilized Petri dish, and place 2 pieces on each piece.
(3) Using a sterilized, fine-pointed inoculation needle or a sewing needle with a handle, take a bit of mold spores (visible to the naked eye) and gently point them on the edge of the agar block, cover the agar block with a coverglass standing next to the slide using sterile tweezers, and cover the dish with a lid.
(4) Add several milliliters of sterile 20% glycerol to the filter paper of the petri dish and stop adding until the filter paper is wet. The petri dish was incubated at 28℃ for a certain period of time, and the slide was removed and observed under a microscope.
3. Cellophane dialysis culture observation
(1) Add 5 ml of sterile water to a mold slant test tube and wash down the spores to make a spore suspension.
(2) Using sterile forceps, cover a sterilized, circular cellophane of the same diameter as the Petri dish with a plate of Chartreuse medium.
(3) Pipette 0.2 ml of spore suspension on the above cellophane plate with a 1 ml sterile pipette and spread well with a sterile glass spatula.
(4) After 48 hours of incubation at 28℃, take out the petri dish, open the lid, separate the cellophane from the medium with tweezers, and then cut a small piece of cellophane with scissors and place it on the slide, and observe it with a microscope.