Experimental principle
The fungus hyphae are coarser, the cells are easily contracted and deformed, and the spores are easily scattered. Therefore, the lactic acid carbonic acid cotton blue staining solution is commonly used in the preparation of the specimen. The mold sample prepared by the dyeing liquid is characterized in that: (a) the cell is not deformed; (b) has a bactericidal and antiseptic effect, and is not easy to dry and can be kept for a long time; (c) the solution itself is blue and has a certain dyeing. effect.
The morphology of the mold under natural growth state is usually observed by a glass slide. This method is inoculated with a mold spore on a suitable medium on a glass slide, and cultured and observed under a microscope. In addition, in order to obtain a clear, intact, and preserved mold form, it can also be observed by a cell paper dialysis culture method. The method utilizes the semipermeable membrane characteristics and light transmittance of the cellophane, and molds the mold on the cellophane covered on the surface of the agar medium, and then cuts a small piece of the cellophane of the long bacteria, and places it on the glass slide and observes it with a microscope.
Experimental reagent
Lactic acid blue cotton staining solution, 20% glycerol, Chad medium plate, potato medium;
Laboratory equipment
Sterile straws, slides, coverslips, U-shaped sticks, scalpels, cellophane, filter paper, etc.
Experimental Materials
Aspergillus sp., Penicillium sp., Rhizopus sp., Mucor sp.;
Experimental procedure
1. General observation
On a clean glass slide, drop a drop of lactic acid carbaceous cotton blue staining solution, use a dissecting needle to take a small amount of spore-forming hyphae from the edge of the mold colony, and then carefully spread the hyphae, then carefully Cover the cover with a cover, taking care not to create air bubbles. Under the microscope, use a low power microscope to observe, and if necessary, change the high power mirror.
2. Slide observation
(1) Put the filter paper slightly smaller than the inner diameter of the bottom of the culture dish into the dish, then put the U-shaped glass rod on it, put a clean glass slide on it, and then tilt the two coverslips on the slide glass respectively. At both ends, cover the lid, put several sets (as needed), stir the culture dish so equipped, pack it with 1.05kg/cm2, sterilize at 121.3 °C for 20 minutes or dry heat sterilization, spare .
(2) Pour 6-7 ml of sterilized potato dextrose medium into a sterilized plate with a diameter of 9 cm. After solidification, cut into a 0.5-1 cm agar block with a sterile scalpel and shovel with a knife tip. Place the agar block on a glass slide in a sterilized Petri dish with 2 pieces on each piece.
(3) Using a sterile needle or a needle with a handle, take a small mold spore (which can be seen by the naked eye), gently spot it on the edge of the agar block, and stand it with a sterile forceps. Cover the slide on the agar block next to the slide and cover the lid.
(4) Add a few milliliters of sterile 20% glycerin to the filter paper of the culture dish, and then add it to the filter paper. After the culture dish was incubated at 28 ° C for a certain period of time, the slide was taken out and observed under a microscope.
3. Cellophane dialysis culture observation method
(1) 5 ml of sterile water was added to the mold beveled test tube, and the spores were washed to prepare a spore suspension.
(2) Cover the sterilized round cellophane with the same diameter and petri dish on the Chad medium plate with sterile forceps.
(3) Pipette 0.2 ml of the spore suspension onto the above cellophane plate with a 1 ml sterile pipette and spread evenly with a sterile glass spatula.
(4) After incubating in a greenhouse at 28 ° C for 48 hours, the culture dish was taken out, the lid was opened, the cellophane was separated from the medium by tweezers, and a small piece of cellophane slide was cut with a pair of scissors, and observed with a microscope.
1. result
The plot illustrates the mold morphology characteristics you have observed.
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