Method of making microscope slides
There are three methods of making microscope slides:
1. Smear method
Smear method is a method of preparation in which the material is spread evenly on the slide.
Smear materials include unicellular organisms, small algae, blood, bacterial cultures, loose tissues of plants and animals, sperm nests, anthers, etc.
Attention should be paid when smearing:
(1) The slide must be clean.
(2) The slide should be held flat.
3) The coating must be uniform. Apply the droplet of coating liquid to the right of the centre of the slide and spread it well with the blade of a dissecting knife or a toothpick, etc.
4) The coating should be thin. Use another slide as a pusher, gently push from right to left along the surface of the slide with the drop of application solution (the angle between the two slides should be 30° to 45°), and apply a uniformly thin layer.
5) Fixation. If fixation is needed, use chemical fixative or desiccation (bacterial) fixation.
6) Staining. Use methylene blue for bacteria and Rachel's stain for blood. The staining solution should cover the entire coated surface.
7) Rinse. Absorb on absorbent paper or bake dry.
8) Seal the film. For long-term storage, use Canadian tree film.
2. Film Pressing Method
Pressure film method is the biological material placed between the slide and cover sheet, applying a certain pressure, the tissue cells will be pressed apart a method of preparation.
(1) take material. Observation of cell division, cell division should be selected exuberant, fresh tissue cells as materials. Such as root tips, stem tip meristematic tissue, bone marrow cells, anthers (pollen mother cells). Sperm nest (spermatogonia) and so on.
(2) Fixation. Material fixation can be determined according to need, immediately after taking the material pressure film observation, can not make a separate fixed treatment (and staining synchronous); take the material is not immediately after the inspection, the material can be fixed with a fixative (generally with alcohol acetate fixative). Fixed 2 ~ 24 hours (depending on the material) after washing with 95% ethanol, preserved in 70% ethanol, standby.
(3) Dissociation. The cells are not easy to disperse the material with hydrolysis separation solution (such as 1N HCl or hydrochloric acid an alcohol solution) treatment, general treatment 6 ~ 20min, after dissociation and rinsing with water before staining.
(4) Staining. There are many kinds of stains. Observation of chromosomes is commonly used to stain with magenta acetate staining solution.
(5) Film pressing. Place the material on a slide, add a drop of water or staining solution, cover the coverslip and gently press the slide with your thumb.
(6) Observation. After pressing the film, it can be observed under the microscope.
