Final answer:
In-vitro staining involves mounting cells on a slide, fixing them, applying stains, and then observing them under a light microscope. Proper preparation of reagents and disinfection of materials is required to ensure a successful staining procedure.
Step-by-step explanation:
The process of in-vitro staining is crucial for examining microscopic specimens and involves several key steps to prepare the sample. Following these steps ensures the sample is properly stained and ready for observation under a microscope.
- Mounting: First, the cells must be smeared on a glass slide. This process is careful work, as the cells must be spread out thinly and evenly across the slide to ensure an accurate stain and easy observation under the microscope.
- Fixing: The smeared cells need to be fixed to the glass slide. This is typically done using chemicals like methyl alcohol or heat to kill and adhere the cells to the slide. Fixation preserves the integrity of cellular structures and prevents degradation over time.
- Staining: Apply the appropriate stain to the slide. The choice of stain depends on the type of specimen and the cellular components of interest. For example, crystal violet may be used for Gram staining of bacteria.
- Observation: After staining, the sample is rinsed and ready to be observed under a light microscope using the oil immersion objective lens. An initial scan is performed to evaluate the overall staining before moving into a more detailed analysis of cellular structures or microorganisms present.
In addition to these steps, ensure all materials, such as forceps, slides, and spreaders, are disinfected with 70% ethanol before use, and that stains are prepared and diluted as required for the specific protocol, such as immunofluorescence, FISH, or hematoxylin and eosin (H&E).