First, the experimental principle Since the affinity between the capsule and the dye is weak and it is difficult to color, the capsule is usually dyed by negative staining, that is, the cells and the background are colored and the capsule is not colored, so that the capsule has a transparent circle around the cells. Since the water content of the capsule is above 90%, it is generally not heated and fixed during dyeing to prevent the capsule from shrinking and deforming. Second, the experimental method The following four staining methods are recommended, among which the wet ink method is simple and suitable for various capsule bacteria. If the phase contrast microscope is used, the effect is better. 1. Negative staining method: (1) Preparation: Take a clean glass slide, add a drop of distilled water, take a small amount of bacteria into the water droplets and mix and coat. (2) Drying: The smear is air-dried or blown dry with a cold air blower. (3) Dyeing: dyeing on the painted surface with red staining solution for 2-3 min. (4) Washing: Wash the red dye solution with water. (5) Drying: The dyed tablets are air-dried or blown dry with a cold air blower. (6) Melanin: Add a small drop of melanin to the left of the stained surface, gently touch the melanin with a smooth glass slide, spread the melanin along the edge of the slide, and then drag it to the right. Melanin becomes a thin layer on the dyed surface and is quickly dried. (7) Microscopic examination: first low magnification, then high magnification observation. Results: The back is gray, the bacteria are red, and the capsule is colorless and transparent. 2, wet ink method (1) Bacterial solution: Add 1 drop of ink to a clean glass slide, pick a small amount of bacteria and mix well. (2) Cover the slide: Put a clean cover slip on the mixture, then put a filter paper on the cover slip and gently press down to absorb the excess bacteria. (3) Microscopic examination: first observe with a low power microscope and then with a high power microscope. RESULTS: The background was gray and the cells were dark, and a bright transparent circle around it was the capsule. 3, dry ink method (1) Bacterial solution: Add 1 drop of 6% glucose solution to one end of the clean glass slide, pick a small amount of Bacillus licheniformis and mix it well, then add 1 ring of ink and mix well. (2) Production: The left hand holds the slide, and the right hand takes a glass slide with a smooth edge. The side of the slide is in contact with the bacterial liquid, so that the bacterial liquid spreads along the contact of the slide, and then at a 30 degree angle. The bacteria liquid is quickly and evenly pulled to one end of the slide to make the bacterial liquid into a film. (3) Drying: It is naturally dry in the air. (4) Fixation: The smear was immersed in methanol, fixed for 1 min, and methanol was immediately decanted. (5) Drying: Above the alcohol lamp, dry with a simmer. (6) Dyeing: dyeing with methyl violet for 1-2 min. (7) Washing: Lightly wash with tap water and dry naturally. (8) Microscopic examination: first observe with a low power microscope and then a high power microscope. Results: The background was gray, the cells were purple, and the capsule showed a clear transparent circle. 4, Tyler method (1) Smear: According to the conventional method, more bacteria can be mixed with water, and the viscous bacteria liquid should be spread as much as possible, but the coated area should not be too large. (2) Drying: Dry naturally in the air. (3) Dyeing: dyeing with Tyler staining solution for 5-7 min. (4) Decolorization: The crystal violet was washed away with a 20% CuSO4 aqueous solution, and the decolorization was moderate (flush 2 times). Drain dry with absorbent paper and immediately add 1-2 drops of cedar oil to the smear to prevent the formation of CuSO4 crystals. (5) Microscopic examination: first observe with a low power microscope and then with a high power microscope. After the observation, pay attention to wipe off the cedar oil on the lens with xylene. Results: The background is blue-violet, the cells are purple, and the capsule is colorless or light purple. Chuangsai Technology provides you with comprehensive professional laboratory reagents. The following promotional consumables are provided by Chuangsai Technology. Welcome to inquire! Item number Product name Brand specification price 1A5105 Standard microscope slide CITOTEST 50 pieces / box Inquiry 188105W Adhesive slide CITOTEST 50 pieces / box Inquiry 10212020C Microscope cover glass 20*20mm CITOTEST 200 pieces / box Inquiry 10212432C Microscope cover glass 24*32mm CITOTEST 200 pieces / box Inquiry 430790 15ml with bottom-end centrifuge tube Corning 50 sticks/package Inquiry 430791 15ml without centrifuge tube Corning 25 sticks/package Inquiry 430828 50ml with bottom-end centrifuge tube Corning 25 sticks/package Inquiry 430829 50ml without centrifuge tube Corning 25 sticks/package Inquiry Third, matters needing attention 1. There should be no air bubbles when you cover the slide, otherwise it will affect the observation. 2. When using the dry ink method, the smear should be placed in a high flame and dried with a simmer, so that the slide does not heat up. 3. 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