What precautions are important when a prepared slide is observed under the microscope?

Answer 1

Microscopes work on the principle of magnifying light rays passing through a tiny object. The object should be transparent or translucent and colored to be properly viewed under a compound microscope. Some of the mandatory requirements to prepare a good slide for viewing are as follows:

Before creating a smear, always check that the slide is clean and perfectly transparent. It should also be microbe free. So, take a slide and wash it first with soap-water, and then wipe it with ethanol (ethyl alcohol). This makes the slide clean and sterilized.

With regard to preparing a slide of bacterial specimen, when you create a smear, do not make a thick layer of smear. Take very little quantity of the inoculum. If by chance you take too much of the inoculum, spread it over the slide to a larger area, such as to avoid a thick smear.

Air drying is necessary, as it lets the bacteria congregate at their places. Heat fix the slide with precision. Too much heat fix can kill the organism, and too little of it will make the organisms too loosely bound to the slide surface. If they are loosely bound, they will fall off when you flood the slide with stain. One good way of identifying the extent of heat fix is to feel it after you pass the slide through the flame. It should neither be too hot nor warm. It should give you the sense of heat but tolerable.

When you stain the slide, do not stain the whole surface of the slide. This wastes much of the stain and is messy. Cleanliness is very important in all science experiments. Just staining the area containing the smear is enough. Usually, stainings are done for a minute or two, but for certain experiments like endospore staining, the extent of staining time may be as long as 10 mins or even more. During such cases, ensure that the stain does not dry over the smear. Maintain liquidity of the stain, as it is to be washed after some time.

While washing the slide after staining, do not let the water stream fall directly on the smear. This may disrupt the smear. Let the stream of water flow slowly along the surface, such that only the stain is flooded and the smear is intact.

While preparing fungal slides, take the stain first, and then the hyphal fragments. Crush the hyphal fragments properly by placing a coverslip over the fragments (avoiding air bubbles) and the slowly tapping it with the butt of a pencil.

Always observe under 10X first. This will give you an idea of the location of a good area for observation. After this, you may prefer to switch over to 45X.

100X objective in compound microscopes is always used as an oil-immersion objective, so do not ever observe at a specimen at 100X without oil.

Answer 2

Do not allow the high power objective to tough the lenses