What is the preparation of normal saline solution?

Why is saline solution used for experiments?

  • Why is saline solution used for in preparation of say cheek cells to be mounted upon a slide and viewed under a microscope?

  • Answer:

    Because if you used pure water the cheek cells would burst due to osmotic pressure. This is called osmolysis. The plasma membrane is permeable to water and the interior of the cell is salty. Pure water would pass through the plasma membrane leading to excess pressure.

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In a wet mount, the specimen is suspended in a drop of liquid (usually water) located between slide and cover glass. The water refractive index of the water improves the image quality and also supports the specimen. In contrast to permanently mounted slides, wet mounts can not be stored over extended time periods, as the water evaporates. For this reason, a wet mount is sometimes also referred to as a “temporary mount” to contrast it from the “permanent mounts”, which can be stored over longer times. The permanently mounted slides use a solidifying mounting medium, which holds the cover glass in place. The naming can be a bit problematic, because it is also possible to make wet mounts that can store over extended time periods. These are special cases, however. Different types of wet mounts: Wet mounts can be made using several different kinds of liquids. Water, immersion oil and glycerin (glycerol) can be used, with water probably being the most commonly used. The source of the water is quite important, especially when observing living specimens. If you use water with a wrong osmotic potential (ie. too much or too little salt and mineral content), then there is the danger of damaging the specimen. A too high salt content can result in the specimen to lose too much water. Too low a salt content, and the specimen may swell and burst. Using water from the natural habitat of the organism: In the case of water organisms, such as algae or ciliates, the liquid water should come directly from the sample. In this case the organism is immersed in its own natural environment. The microscopist uses a dropper to place a drop of pond water directly on the microscope slide. Using 0.9% salt water: In some cases water from the natural habitat may not be available. This is the case when observing bacteria or molds grown on petri-dishes. Yoghurt bacteria, for example, need to be diluted a lot before being able to observe them, otherwise they are too dense to be observed as single cells. In this case it is necessary to mix some salt (NaCl) into some water to ensure an optimal osmotic potential. This “physiological saline”, as it is called, can be made by dissolving 9 grams of table salt (NaCl) in 1 liter of water (or 0.9g Nacl in 100ml of water). Using tap water: If one wants to observe non-living specimens, such as dust samples, sand grains, or thin section cuts of plant material, then it is also possible to use regular tap water. These specimens are not osmotically sensitive. If the specimen is observed without water, in a dry condition, then the resolution and image quality may not be sufficiently high. I advise you to try out both to see the difference. The following post includes images of pollen grains mounted in air and water, for comparison: The effect of the mounting medium on specimen and image quality Using immersion oil: Some wet mounts are not made with water, but by using immersion oil. Immersion oil is usually placed on top of the cover glass. In this case the specimen does not get into contact with the oil. It is also possible to submerge the specimen in the oil, however. Heat-fixed bacteria can be observed directly by placing a drop of immersion oil on the specimen, without cover glass. The oil-immersion objective is then rotated directly into the oil for observation. It goes without saying, that this procedure can only be used for specimens that do not contain water (and are, therefore, not living). It also only works for specimens that stick to the glass slide – there is no cover glas. If you need to observe these specimens with a lower magnification (ie. no immersion objective), then one needs to use a cover glass, of course. Other specimens, such as synthetic textile fibers, are hydrophobic in nature, and do not like to be mixed with water. They tend to float on top of the water drop and this can be cause for air bubbles. In this case I also recommend to use immersion oil and a cover glass to keep the sample flat. Pure glycerin or glycerin-water mixtures: Glycerin has a strong tendency to withdraw water from the sample. For this reason it also acts as a preservative. On the down side, the glycerin may therefore cause the specimen to shrink and deform. Especially algae and other water organisms are sensitive to dehydration. Other specimens, such as sectioned or microtomed plant material are not as sensitive. The reason why glycerin is used is because of its high refractive index. This may be necessary to see certain structures. If a lower refractive index is needed, then one should mix some water into the glycerin. It is possible to seal the glycerin mount by applying nail polish to the sides of the cover glass. This will hold the cover glass in place for longer time periods. This is then an example of a wet mount, which was made into a permanent mount.

John

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