The preparation method of microscope slides
1. Smear method
Smearing method is a method of preparing slides by uniformly coating materials onto them.
Smear materials include single-cell organisms, small algae, blood, bacterial culture media, loose tissues of animals and plants, testes, anthers, etc.
When smearing, attention should be paid to:
(1) The glass slide must be cleaned.
(2) The glass slide should be flat.
(3) The coating must be uniform. Apply the droplet to the middle right of the glass slide and spread it evenly with a dissecting blade or toothpick.
(4) The coating should be thin. Using another glass slide as a slide, gently push from right to left along the surface of the glass slide with the applied liquid (the angle between the two slides should be 30 ° to 45 °), and apply a uniform thin layer.
(5) Fixed. If fixation is required, chemical fixatives or drying methods (bacteria) can be used for fixation.
(6) Staining. Bacteria use methylene blue, and blood uses Wright staining solution. The staining solution should cover the entire coating surface.
(7) Rinse. Use absorbent paper to absorb or dry.
(8) Seal the film. Long term preservation using Canadian tree film.
2. Tablet pressing method
The compression method is a method of slicing biological materials by placing them between a glass slide and a cover, applying a certain pressure to disperse tissue cells.
The general process of tablet pressing method:
(1) Material selection. To observe cell division, fresh tissue cells with vigorous cell division should be selected as materials. Such as root tip, stem tip meristem, bone marrow cells, anthers (pollen mother cells). Sperm nests (spermatocytes), etc.
(2) Fixed. The material fixation can be determined according to the needs, and the sample should be immediately compressed and observed. It is not necessary to perform separate fixation treatment (synchronous with staining); If the material is not inspected immediately after sampling, it can be fixed with a fixative (usually acetic acid alcohol fixative). After fixing for 2-24 hours (depending on the material), rinse with 95% ethanol and store in 70% ethanol for later use.
(3) Separation. Materials that are not easily dispersed by cells should be treated with a hydrolysis separation solution (such as 1N HCl or hydrochloric acid semen) for 6-20 minutes. After dissociation, they should be rinsed with water before staining.
(4) Staining. There are many types of dyes. Chromosomes are commonly stained with acetic acid magenta staining solution.
(5) Press the tablet. Place the material on a glass slide, add a drop of water or dye solution, cover the slide with a cover, and gently press it with your thumb.
(6) Observe. After compression, it can be observed under a microscope.
