For a non-medical student, the variety of microscopes that are available in the market may be confusing. This is because there are so many kinds of microscope that are built different purposes. There are monocular microscopes, compound light microscopes, low power stereoscopic microscopes, binocular microscopes, forensic comparison microscopes, and even metallurgical microscopes. As its name, implies, the uses of microscopes can range from verifying the authenticity of jewelries to examining anti-bodies. It can be frustrating to browse through this selection when you want to choose the kind of microscope that will fit your needs.

Most of the microscopes stated above are fluorescence microscopes. And in order to understand why epifluorescence microscope is so effective, it is important to look its functions. When you are using fluorescence microscopy, the light source comes from the sample itself. Fluorescence microscopy is based upon the concept that there are certain materials that emits energy which can be detected as a visible light. This detection is possible when it is irradiated with a specific light wavelength. The sample being used can either be treated with some fluorescing substances or it can be fluorescing on its original form.

Basics about Fluorescence Microscopes

Fluorescence microscopes are mainly used for the study of microscope elements that cannot be seen in the naked eye. Unlike the conventional microscopes though, the fluorescence microscope is used on specimens that emit fluorescence light themselves. Usually, a researcher would need to put fluorescing dyes such as fluorochromes so that the sample can be seen with a high microscopic contrast.

As you can observe, the basic of why fluorescence microscopy is used is that the optics it uses provides a method of high-quality illumination of the sample. In addition, you should also note that filters are mainly used in fluorescence microscopes so that the appropriate wavelength of emission and excitation will be achieved. The excitation filters are the ones responsible for setting a limited range of wavelength that is required. These barrier filters will separate the unabsorbed exciting light from the emitted light so that it can produce the best result possible.

Difference between Dark Ground Condenser and Epifluorescence

There are basically two types of illumination available in fluorescence microscopy. The first is the dark-ground condenser which just facilitates the separation of the exciting light and the fluorescence light. With the introduction of epifluorescence though, it is now possible for the exciting light to reach the preparation level with the use of a dichroic mirror and condenser.

Functions of Epifluorescence Microscope

So the basic task of an epifluorescence microscope is to get the sample get excited. To do this, the microscope has to sort out the excitation light that comes from the sample from the weaker light from the light emission. The epifluorescence microscope needs to filter the radiation so that only the desired wavelength would be retained. This succeeding radiation will then collide with specimen atoms when as they get excited to a higher level of energy so when they relax at a lower energy level, they will be able to emit light.

In order to become visible though, the emitted light from the microscope must be separated from the excitation light at the second filter. In this area, the lower energy of emitted light with a longer wavelength is used. So the fluorescing areas will be observed with a high contrast against the dark background on the microscope.

Because of the key features of an epifluorescence microscope, it is now becoming rapidly used in the biological and medical field. This microscopy technique has made it possible for people to determine and identify the cellular components of each sample with a high amount of specificity. For example, impurities and disease conditions can now be studied effectively by using epifluorescence microscopy.

People who use Epifluorescence Microscopes

So to answer the question, who uses epifluorescence microscope? The answer is a lot of people involved in different fields make use of it. However, it is observed that the primary consumers of the epifluorescence microscope are the medicine industry. Diagnostic test has also been developed by using the monoclonal antibodies wherein green, blue, and red fluorescence dyes are attached to reveal the various components found in the sample.

Conclusion

It is undeniable that the epifluorescence microscopy technology available today has made developments in science and technology develops at a much faster rate. With the availability of the epifluorescent technology, cures and anti-bodies for various diseases can be discovered quicker than ever before. Because of this, we can truly say that it is important for medical students to learn how epifluorescence microscopes are used because this is the way to cellular biology progress for the future. Read more on this subject