First, the basic principle The so-called single staining method is a method of dyeing bacteria with a single dye. This method is easy to operate and is suitable for the observation of the general morphology of the bacteria. Phyllanthus Emblica L,Extract Of Fructus Cynanchum,Phyllanthus Emblica Extract,Rhizoma Glycyrrhizae Extract NANJING ZELANG MEDICAL TECHONOLOGY CO., LTD , https://www.hawfitness.com
In neutral, alkaline or weakly acidic solutions, bacterial cells are usually negatively charged, so basic dyes are commonly used for dyeing. A basic dye is not a base. Like other dyes, it is a salt. When ionized, the dye ions are positively charged and easily combine with negatively charged bacteria to color the bacteria. For example, methylene blue (methylene blue) is actually methyleneblue chloride (abbreviated as MBC), which can be ionized into positive and negative ions:
MBC→methylene blue++chloride-
Positively charged dye ions can stain bacterial cells blue. In addition to methylene blue, commonly used basic dyes include crystal violet, basic fu-chsin, and red (also known as safranine).
Bacteria are small in size and relatively transparent. They are often difficult to identify without staining, and after coloration, they are in sharp contrast with the background, making it easy to observe under a microscope.
Second, equipment microscope, alcohol lamp, glass slide, inoculating ring, double-layer bottle, lens paper, saline;
Lu's alkaline blue dyeing solution, carbolic acid red dyeing solution;
Staphylococcus aureus, Bacillus subtilis.
Third, the operation steps
1. Smear Take two clean slides, each drop of a small drop of normal saline in the center of the slide, and use aseptic operation (Fig. IV-1, specific operation, refer to experiment 1) to pick up Staphylococcus aureus and dry grass respectively. Bacillus was placed in water droplets of two slides (one for each strain), homogenized and coated into a film. Note that the saline solution should not be too much, and the smear must be uniform.
2. Dry and dry naturally at room temperature.
3. Fix the smear face up and pass the 2-3 times on the flame to solidify the cytoplasm to fix the bacteria and make it not easy to fall off. But you can't bake on the flame, or the bacteria will be destroyed.
4. Dyeing Place the specimen in a horizontal position and add the staining solution to the smear film. The length of the dyeing time depends on the dyeing solution. Lu's alkaline blue dyeing solution is dyed for about 2-3 minutes, and the carbolic acid red dyeing solution is dyed for about 1-2 minutes.
5. Washing After the dyeing time is reached, rinse with tap water until the water that is washed is colorless. Note that the flushing water flow should not be too fast, too large, and the water flows down from the upper end of the slide to avoid direct rushing at the smear. After rinsing, dry the specimen or blow it dry with a hair dryer. After it is completely dry, it can be observed under the oil microscope.
6. Microscopic examination was performed under the microscope according to the experimental procedure of Experiment 2.
Fourth, the experimental report