What is immersion oil

However, once you use the x objective lens, the light refraction when using a dry lens is noticeable. If you can reduce the amount of light refraction, more light passing through the microscope slide will be directed through the very narrow diameter of a higher power objective lens. By placing a substance such as immersion oil with a refractive index equal to that of the glass slide in the space filled with air, more light is directed through the objective and a clearer image is observed.

Take a look at the images captured below. The images were captured using the UX1 microscope with the x Achromat objective lens and the x Plan Achromat objective lens. Observation was performed dry and with immersion oil. Notice the difference in image quality between the images captured dry versus those captured with immersion oil.

Microscopy image of duodenum captured using x Achromat objective lens, dry. If you have any questions about using immersion oil with your microscope, contact Microscope World.

The specifications are the following:. Here are some of the common types of immersion oils based on viscosity and applications:. The blog on chemicals. The blog, its authors and affiliates accept no responsibility for any accident, injury or damage caused in part or directly from following the information provided on this website. We do not recommend using any chemical without first consulting the Material Safety Data Sheet which can be obtained from the manufacturer and following the safety advice and precautions on the product label.

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You can then look down the eyepieces again. Use only the fine focus to adjust the field of view. Although high power objectives have a spring-loaded nose, coarse focussing at this stage can easily result in cracking the cover glass or slide and can also damage the objective front lens.

Even if you plan to examine other slides, you should remove the oil from the objective at this stage to prevent possible contamination of other parts of the microscope. Immersion oil can and will penetrate and damage microscope components and objectives not suited for immersion. Remove excess oil using a lens cleaning tissue with a single sweep across the lens.

Keep wiping the objective front lens with a clean piece of tissue for each wipe until no trace of oil remains. Commercial oil removal solutions are available or a small amount of xylene can be used to finally clean the lens. Again, it is important to check the recommendations from the objective manufacturer before applying any solutions to the lens. The water immersion objective is highly recommended when imaging live cells which are in cell medium.

There are two types of water immersions objectives s. The dipping objectives are manufactured to provide a very long working distance. They are also manufactured with steeply angled nose-pieces which are constructed from inert material such as ceramic. Water immersion objectives are used in a manner similar to oil immersion objectives, but with water in place of the drop of oil.

One of the advantages of using a water immersion objective is simply that water is used as the immersion medium. This is obviously easy to apply and clean off. Additionally, you do not need to use specific immersion oil depending on the imaging you are carrying out, nor do you need to use an immersion medium as specified by the manufacturer of the microscope and objective. There are, however, some disadvantages when using a water immersion objective.

A higher resolution is achievable with oil immersion objectives compared to the aqueous objectives. In addition, due to the viscosity of water compared to oil , the use of water immersion objectives can be susceptible to vibrations and small air movements. Such artefacts can be overcome by ensuring the microscope is placed on an anti-vibration table.

Alternatively, a more simple solution is to place a special ring which sits over the slide to create a small pool of water. The final disadvantage of water immersion objectives is their cost. Some water immersion objectives can cost as much as a complete research grade microscope. Leica offers a Water Immersion Micro Dispenser which overcomes the potential problem of water evaporation during long-term live-cell imaging or screening experiments s.

Figure 6. Overall, the main advantage of water immersion objectives comes when imaging live cells and tissue. This is mainly due to the fact that oil immersion objectives are not suited for imaging through the cell or tissue chambers used for live cell microscopy.

Living cells are usually contained within a chamber as well as being covered with cell medium or buffer. The chamber and the medium help to ensure that the cells or tissue are maintained within a stable environment whilst imaging. As a consequence, the optimum focal distance will be at a relatively large distance from the cover of the chamber. Furthermore, using an oil immersion objective to view cells within an aqueous medium would add additional refraction problems as oil and water have different refractive indices.

When viewing living cells within a chamber, the light path will encounter different refractive indices. The light which forms the specimen image will be refracted at each stage which can lead to spherical aberrations. Despite the refraction which can occur at the water, glass or plastic interfaces, water immersion objectives are usually corrected for this. In addition, some water immersion objectives have correction collars.

These rings around the objective barrel can be adjusted to suit the different thickness of cover glasses.